Validation of an extraction method for the quantification of soluble free and insoluble bound phenolic compounds in wheat by HPLC-DAD

Wheat is one of the three most widely grown grain crops and the most consumed in Europe. Wheat contains different phenolic compounds, mainly phenolic acids and flavonoids that are not evenly distributed in the wheat, being most abundant in the bran. These phenolic compounds are antioxidants against free radicals and other reactive oxygen species. Therefore, over the last few decades, there is a great interest to study and quantify these compounds present in wheat.This work proposes a HPLC-DAD method that allows a good separation of different phenolic compounds occurring as soluble free and insoluble bound compounds in wheat brand and flour. The extraction and the chromatographic conditions allowed the characterization of forty-two phenolic compounds, among phenolic acids (HBA, HCA, DFA and TFA), phenolic aldehydes, phenolic alcohols and flavonoids (flavones). This method is simple, sensitive, reproducible and accurate, and only a HPLC-DAD equipment is required. Thirty-four soluble and insoluble phenolic compounds can be quantified in wheat flour and brand. Hydroxytyrosol, a potent antioxidant compound, has been identified and quantified in wheat for the first time.     

Development of a reverse-transcription loop-mediated isothermal amplification assay to detect avian influenza viruses in clinical specimens

In recent years, the avian influenza has brought not only serious economic loss to the poultry industry in China but also a serious threat to human health because of the avian influenza virus(AIV) gene recombination and reassortment. Until now, traditional RT-PCR, fluorescence RT-PCR and virus isolation identification have been developed and utilized to detect AIV, but these methods require high-level instruments and experimental conditions, not suitable for the rapid detection in field and farms. In order to develop a rapid, sensitive and practical method to detect and identify AIV subtypes, 4 specific primers to the conserved region of AIV M gene were designed and a loop-mediated isothermal amplification(RT-LAMP) method was established. Using this method, the M gene of H1–H16 subtypes of AIV were amplified in 30 min with a water bath and all 16 H subtypes of AIV were able to be visually identified in presence of fluorescein, without cross reaction with other susceptible avian viruses. In addition, the detection limit of the common H1, H5, H7, and H9 AIV subtypes with the RT-LAMP method was 0.1 PFU(plaque-forming unit), which was 10 times more sensitive than that using the routine RT-PCR. Further comparative tests found that the positivity rate of RT-LAMP on detecting clinical samples was 4.18%(14/335) comparing with 3.58%(12/335) from real-time RT-PCR. All these results suggested that the RT-LAMP method can specifically detect and identify AIV with high sensitivity and can be considered as a fast, convenient and practical method for the clinic test and epidemiological investigation of AIV.     

西藏引进良种奶牛高原适应性观察与研究

文章旨在研究拟通过测定引进奶牛进入高原后疾病发生率、生产性能发挥情况及血液主要生理指标变化,继而综合评价引进奶牛高原适应性。试验选取从澳大利亚引进的纯种娟姗牛和荷斯坦奶牛,两个品种奶牛均分为高山病发病组、生理指标组、生产性能组,每组均包含对照组和试验组。结果表明,高山发病组:引入一年后,荷斯坦奶牛因高山病死亡率为26.5%,娟姗牛因高山病死亡率为8%,娟姗牛高山病发病率及死亡率显著低于荷斯坦奶牛,且氧气浓度与高山病发病率密切相关;生理指标组:与奶牛引入前组(Before introduction,BID组)相比,奶牛引入前组(After introduction,AID组)娟姗牛和荷斯坦奶牛进入高原1年后,红细胞(RBC)、血红蛋白(HGB)极显著提高(P <0.01),红细胞平均体积(MCV)显著提高(P <0.05),平均血红蛋白浓度(MCHC)极显著降低(P <0.01),此外呼吸、脉搏均有提高;生产性能组:与奶牛引入前组(Before introduction,BID组)相比,奶牛引入前组(After introduction,AID组)娟姗牛和荷斯坦奶牛进入高原后,产奶量、体细胞数极显著降低(P <0.01),乳蛋白率显著降低(P <0.05),此外犊牛初生重较低,且死亡率明显高于对照组。由此可见,引进奶牛无论是疾病抵抗还是生产性能发挥方面均未表现出对高原环境的长期适应性。     

啶酰菌胺对黄瓜灰霉病防治效果的综合评价

利用菌丝生长速率法检测了6株灰葡萄孢(Botrytis cinerea)对6种药剂的抗性背景,发现它们对啶酰菌胺具有不同水平的抗性。同时结合盆栽试验和田间试验评价了50%啶酰菌胺水分散粒剂对黄瓜灰霉病的防治效果。盆栽试验中,50%啶酰菌胺水分散粒剂对具有低等抗性的2株灰葡萄孢的防治效果分别为53.49%和53.04%,对3株高等抗性的灰葡萄孢的防治效果分别为50.16%、51.67%和32.24%。说明灰葡萄孢的抗性发展已经严重影响了啶酰菌胺的杀菌活性。2013年和2014年的田间试验中,50%啶酰菌胺水分散粒剂施药量为375g·(667m2)-1时,防治效果分别为94.34%和90.39%。在施药量为300g·(667m2)-1时,50%啶酰菌胺水分散粒剂与对照药剂50%腐霉利可湿性粉剂的防治效果都很显著。综上所述,现阶段啶酰菌胺对黄瓜灰霉病仍具有很好的防治效果。     

玉米骨干亲本黄早四抗病基因遗传传递规律的初步研究

对玉米抗病基因SSR标记的分析表明,在黄早四、4个黄早四衍生系和衍生系配制的4个杂交种中都存在矮花叶病抗性基因mdml(t)与玉米茎腐病抗性基因Rfg1,但不具有茎腐病抗性基因Rpi1。鲁单981具有南方锈病抗性基因RppQ,其余8个材料都不含该基因。对小斑病抗性基因STS标记的扩增结果显示9,个材料具有大小相同的带型,序列分析显示黄早四具有抗性基因rhm。SSR标记和STS标记分析结果同田间抗性相符合,两种分子标记可有效地用于玉米抗性基因的遗传传递分析。     

甜樱桃品种SSR指纹检索系统的开发及遗传多样性分析

用SSR技术开发了甜樱桃( Prunus avium ) 指纹检索系统并进行遗传多样性分析。18对樱桃、桃和杏的引物在19份甜樱桃及2份草原樱桃( P. fruticosa) 品种中共扩增出83个等位位点, 每个 SSR位点的等位位点数2 ～8 个, 平均416 个, 多态性信息量( PIC) 变化范围为0.38 ～0.80, 平均为 0.64。7个甜樱桃品种具有特殊位点或特殊带型。利用UDP98-414、UDP98-406、UDP96-001及PMS40等4 对引物开发的指纹检索系统, 可以区分18个甜樱桃品种。根据遗传距离进行聚类分析, 19个甜樱桃品种 分成2组, 甜樱桃品种间的聚类结果基本反映了供试材料之间的亲缘关系。     

玉米花粉导入外源DNA研究

为探讨花粉管导入外源DNA方法在玉米育种中的应用效果,以早D502玉米自交系、78599玉米自交系及大豆（丹304）为DNA供体,分别采用柱头涂抹法、开苞导入法、花粉粒浸泡法进行转化,比较3种方法导入后的结实率及变异率。结果表明：开苞导入法结实率最高,其变异率仅为1.4%;花粉粒浸泡法的结实率和变异率均最低;柱头涂抹法结实率较低,但变异率较高。2007年和2009年的导入结果显示,将花粉管导入外源NDA技术作为常规育种辅助手段是完全可行的,对促进植物基因工程与常规育种的有机结合具有现实意义。     

Application of mouse model for evaluation of recombinant LpxC and GmhA as novel antigenic vaccine candidates of Glaesserella parasuis serotype 13

Glaesserella parasuis (G. parasuis) has been one of the bacteria affecting the large-scale swine industry. Lack of an effective vaccine has limited control of the disease, which has an effect on prevalence. In order to improve the cross-protection of vaccines, development on subunit vaccines has become a hot spot. In this study, we firstly cloned the lpxC and gmhA genes from G. parasuis serotype 13 isolates, and expressed and purified their proteins. The results showed that LpxC and GmhA can stimulate mice to produce IgG antibodies. Through testing the cytokine levels of interleukin 4 (IL-4), IL-10 and interferon-γ (IFN-γ), it is found that recombinant GmhA, the mixed LpxC and GmhA can stimulate the body to produce Th1 and Th2 immune responses, while recombinant LpxC and inactivated bacteria can only produce Th2 immune responses. On the protection rate for mice, recombinant LpxC, GmhA and the mixture of LpxC and GmhA can provide 50%, 50% and 60% protection for lethal dose of G. parasuis infection, respectively. The partial protection achieved by the recombinant LpxC and GmhA supports their potential as novel vaccine candidate antigens against G. parasuis.     

种植密度对适宜机械化栽培高粱品种产量及生理特性的影响

以适宜机械化种植的高粱品种辽杂37和晋杂34为试验材料,分别对6个密度处理的产量、产量构成因素、叶面积指数、叶绿素含量、光合速率及群体透光率进行了测定与分析。结果表明：辽杂37和晋杂34均在密度为13.5万株/hm ²时产量最高,分别可达10 551.0和10 324.5kg/hm ²。其中辽杂37在12.0万~15.0万株/hm ²、晋杂34在12.0万~13.5万株/hm ²密度时穗数、穗粒数和穗粒重协调效果较好,可保持较高产量水平。当密度达到15.0万株/hm ²时,随着密度的继续增加将导致群体光合速率和透光率大幅度下降,影响干物质积累和产量形成。     

The solid-solution distribution of copper added to soils: influencing factors and models

Purpose

A series of empirical and mechanistic geochemical models were developed to describe the solid-solution partitioning of copper (Cu) in typical fresh spiked Chinese soils.

Materials and methods

The influence of soil properties on Cu partitioning was assessed in a wide range of soils using multiple regression analysis. Geochemical models (WHAM VI and Visual MINTEQ) and simulation analyses in combination with experimental data (i.e., the bulk of soil properties and Cu contents) were performed in order to provide additional insight into the mechanisms controlling the Cu partitioning. Calculation of soluble Cu contents based on the two models was then simplified and optimized by adjusting input variables, and the calibrated outputs were used to produce reasonable predictions of soluble metal concentrations.

Results and discussion

The results of the multiple regression analyses presented in this paper show strong correlations between soluble Cu concentrations and soil Cu concentrations and properties, with adjusted coefficients of determination (R_adj²) ranging between 0.84 and 0.91. Soil organic carbon (OC) content was an insignificant factor in most cases, but the active fraction of dissolved organic matter was important in improving model estimates. The best fit of root mean square error (RMSE) varied between 0.42 and 0.77 for the WHAM VI model and between 0.28 and 0.57 for the Visual MINTEQ model across all pH categories.

Conclusions

The models presented in this paper are suitable for investigating and simulating Cu solid-solution partitioning in a wide range of Chinese soils.