Development of Nucleic Acid Lateral Flow Immunoassay Detection Method for Foot-and-mouth Disease Virus

In this article, through the combination of nucleic acid probes and immune chromatography, a simple, sensitive and specific detection system——nucleic acid lateral flow immunoassay (NALFIA) for amplifing foot-and-mouth disease virus (FMDV) 3D RT-PCR products was established.An ultrasensitive nucleic acid biosensor (NAB) based on streptavidin-labeled gold nanoparticles dual labels and lateral flow strip biosensor (LFSB) were used in this system.The biotinylated goat anti-rabbit IgG was marked to the NC membrane as the alleged strip and the anti-digoxin antibody was labeled to the NC membrane to capture the digoxin probe.After assemblying gold-labeled strip and detecting RT-PCR products, the detection limit of NALFIA was 0.3×10^-3 to 3×10^-3 μg/μL.The NALFIA was compared with agar gel electrophoresis analysis, the results showed that the sensitivity of NALFIA was higher than agar gel electrophoresis.There was an excellent agreement between the two methods.NALFIA was a method with high sensitive, low cost and short time.In conclusion, this method provided a good alternative to detect FMDV.     

3,5-二硝基水杨酸法与酶法测定甜玉米还原糖和蔗糖含量的比较

针对目前测定甜玉米还原糖和蔗糖含量的3,5-二硝基水杨酸法和酶法进行比较研究,分析两者在育种中的应用。结果显示:酶法的测量准确度比3,5-二硝基水杨酸法高,且其稳定性和重现性都优于3,5-二硝基水杨酸法。通过两种方法对10个甜玉米杂交种种子的还原糖和蔗糖含量的测定,证实了两种方法在甜玉米还原糖和蔗糖含量的检测上是一致的。从育种实践考虑,酶法适于样品数较小的检测,3,5-二硝基水杨酸法更适合于大批量样品的检测。     

利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.     

Whole-genome sequence of a novel Chinese cyprinid herpesvirus 3 isolate reveals the existence of a distinct European genotype in East Asia

Cyprinid herpesvirus 3 (CyHV3), also known as koi herpesvirus (KHV), can be subdivided primarily into European and Asian genotypes, which are represented by CyHV3-U or CyHV3-I and CyHV3-J, respectively. In this study, the whole genome sequence of a novel Chinese CyHV3 isolate (GZ11) was determined and annotated. CyHV3-GZ11 genome was found to contain 295,119 nucleotides with 52.9% G/C content, which is highly similar to those of published CyHV3-U, CyHV3-I, and CyHV3-J strains. With reference to CyHV3-U, CyHV3-I, and CyHV3-J, CyHV3-GZ11 was also classified into 164 open reading frames (ORF), which include eight repeated ORFs. On the basis of the 12 alloherpeviruses core genes, results from phylogenetic analysis showed that CyHV3-GZ11 had closer evolutionary relationships with CyHV3-U and CyHV3-I than with CyHV3/KHV-J, which were also supported by genome wide-based single nucleotide substitution analysis and the use of a series of developed molecular markers. This study was the first to reveal the presence of a distinct European CyHV3 genotype in East and Southeast Asia at a whole genome level, which will evoke new insights on exploring the origin, evolution, and epidemiology of the virus.     

Inlfuence of FSH Treatment on Expression of CDC25A,TSSK3 and P53in Vitro Cultured Sertoli Cells of Calf

CDC25A, TSSK3 and P53 expressionsin vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of...     

How does water and salt stress affect the germination and initial growth of Brazilian soya bean cultivars?

The effects of water and salt stress on rate of germination and seedling growth were investigated under laboratory conditions in 46 soya bean genotypes from Central-West region of Brazil to verify how these stresses may limit crop establishment during the initial growth stage and also to identify the most tolerant genotypes to drought and salinity. Mild water and salt stresses were imposed by seed exposure to –0.20 MPa iso-osmotic solutions with polyethylene glycol—PEG 6000 (119.57 g/L) or NaCl (2.357 g/L) for 12 days at 25°C. The germination percentage, seedling length and seedling dry matter were measured, and then, salt or drought tolerance indexes were calculated. The “NS 5909 RG,” “NS 7000 IPRO,” “NS 7338IPRO,” “FPS Solimões RR,” “NS 5151 IPRO,” “SYN 13610 IPRO,” “LG 60177 IPRO,” “NS 6909 IPRO” and “BMX Desafio RR” were identified as the most drought-tolerant genotypes, whereas under salinity conditions, the genotypes “5D 615 RR,” “BMX Desafio RR,” “5D 6215 IPRO” and “BMX Ponta IPRO” were identified as tolerant. The “BMX Desafio RR” is the genotype most adapted to both stress conditions and, therefore, should be used under conditions of water shortage and excess salt in the soil at sowing time.     

HMGCR基因多态性与肉牛屠宰性状相关性分析

MHGCR是胆固醇生物合成过程中的限速酶,在内源胆固醇生物合成过程中起着重要的作用。研究以牛HMGCR基因为候选基因,采用PCR-SSCP和测序技术在利杂牛、利西杂牛、秦川牛、西杂牛和夏西杂牛五个群体共123个个体中检测遗传变异情况。结果表明,检测到AA和AB共2种基因型,没有发现BB基因型,测序分析发现一处SNP多态位点(GenBank:BC153262.1 11137TC)。不同基因型与屠宰性状的相关性分析显示,在利西杂群体中AA基因型个体与AB基因型个体相比,AA基因型个体的屠宰率和牛柳指标达到了极显著水平(P0.01),而AA基因型的上脑重显著高于AB基因型(P0.05),其余各项指标基因型间差异不显著(P0.05)。因此,HMGCR基因单核苷酸多态性对利西杂牛脂肪沉积有一定的影响,可作为影响屠宰性状的候选基因,指导肉牛的育种工作。     

水分胁迫下转DREB3基因抗旱大豆的生理反应

采用田间试验方法,在苗期和初花期自然状态、水分胁迫以及充足灌水条件下,研究了转DREB3基因抗旱大豆叶片、茎、根中超氧化物歧化酶(SOD)活性,丙二醛(MDA)、脯氨酸和蔗糖含量的变化。结果表明,在自然状态下,苗期水分胁迫第5天时,转DREB3基因抗旱大豆叶片SOD活性显著高于受体大豆东农50,初花期无明显变化;苗期和初花期转DREB3基因抗旱大豆叶片、茎和根中丙二醛、脯氨酸含量与受体大豆东农50相比无明显差异;苗期转DREB3基因抗旱大豆叶片和根中蔗糖含量显著低于受体大豆东农50,初花期差异不显著。在水分胁迫条件下,转DREB3基因抗旱大豆SOD活性显著高于受体大豆东农50,丙二醛含量显著低于受体大豆东农50,初花期水分胁迫第10天时叶片和茎中脯氨酸含量显著高于受体大豆东农50,蔗糖含量显著低于受体大豆东农50。充足灌水管理下,转DREB3基因抗旱大豆与受体大豆东农50相比,苗期和初花期叶片、茎和根中SOD活性、丙二醛和脯氨酸含量无明显差异;苗期转DREB3基因抗旱大豆叶片中蔗糖含量显著低于受体大豆东农50,但在初花期水分胁迫10天时转DREB3基因抗旱大豆叶片中蔗糖含量显著高于受体大豆东农50,苗期和初花期茎和根中蔗糖含量无显著差异。