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.     

Influences of sand cover on erosion processes of loess slopes based on rainfall simulation experiments

Aeolian-fluvial interplay erosion regions are subject to intense soil erosion and are of particular concern in loess areas of northwestern China.Understanding the composition,distribution,and transport processes of eroded sediments in these regions is of considerable scientific significance for controlling soil erosion.In this study,based on laboratory rainfall simulation experiments,we analyzed rainfall-induced erosion processes on sand-covered loess slopes(SS)with different sand cover patterns(including length and thickness)and uncovered loess slopes(LS)to investigate the influences of sand cover on erosion processes of loess slopes in case regions of aeolian-fluvial erosion.The grain-size curves of eroded sediments were fitted using the Weibull function.Compositions of eroded sediments under different sand cover patterns and rainfall intensities were analyzed to explore sediment transport modes of SS.The influences of sand cover amount and pattern on erosion processes of loess slopes were also discussed.The results show that sand cover on loess slopes influences the proportion of loess erosion and that the compositions of eroded sediments vary between SS and LS.Sand cover on loess slopes transforms silt erosion into sand erosion by reducing splash erosion and changing the rainfall-induced erosion processes.The percentage of eroded sand from SS in the early stage of runoff and sediment generation is always higher than that in the late stage.Sand cover on loess slopes aggravates loess erosion,not only by adding sand as additional eroded sediments but also by increasing the amount of eroded loess,compared with the loess slopes without sand cover.The influence of sand cover pattern on runoff yield and the amount of eroded sediments is larger than that of sand cover amount.Furthermore,given the same sand cover pattern,a thicker sand cover could increase sand erosion while a thinner sand cover could aggravate loess erosion.This difference explains the existence of intense erosion on slopes that are thinly covered with sand in regions where aeolian erosion and fluvial erosion interact.     

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...     

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。     

An improved method for extraction of sorghum polymeric protein complexes

A method for fractionating sorghum proteins using extraction solvents and techniques designed to obtain polymeric protein structures (especially disulfide linked) was developed. Extraction and separation conditions were optimized in terms of completeness of protein extraction, sample stability, and analytical resolution. After pre-extraction of albumins and globulins, a 3-step sequential procedure involving no reducing agents was applied to ground whole sorghum flour. The three fractions obtained represented proportionally different protein polymer contents and molecular weight distribution as evidenced by comparative size exclusion chromatography. Protein composition also varied among the extracts with differences in kafirin composition and non-kafirin proteins detected in the fractions by RP-HPLC and SDS-PAGE analysis. The ability to quantify and further characterize sorghum polymeric protein complexes will be useful for additional studies linking protein structures with functionality and digestibility and variations for these properties within diverse sorghum germplasm.