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.     

Expression and mechanisms of hsa_circ_087631 in patients with primary biliary cholangitis

AIM To explore the expression and mechanisms of circular RNA hsa_circ_087631 in the patients with primary biliary cholangitis （PBC）. METHODS RT-qPCR was used to detect the expression of hsa_circ_087631 in PBC patients and healthy controls. Hsa-miR-346-overexpressing lentiviral vector pLenti-EF1a-EGFP-F2A-Puro-CMV-MCS was constructed and transfected into human acute T cell leukemia Jurkat cells， and then the expression levels of hsa_circ_087631， Bcl-6 mRNA and interleukin-21 （IL-21） mRNA were detected by RT-qPCR. Dual-luciferase reporter assay was performed to identify the interactions between hsa_circ_087631 and hsa-miR-346. RESULTS The expression of hsa_circ_087631 in the PBC patients was significantly increased compared with the healthy subjects. Hsa-miR-346-overexpressing lentiviral vector pLenti-EF1a-EGFP-F2A-Puro-CMV-MCS was successfully constructed. The expression of hsa-miR-346 was significantly increased after the hsa-miR-346-overexpressing lentiviral vector was transfected into the Jurkat cells， while the expression levels of hsa_circ_0087631， Bcl-6 mRNA and IL-21 mRNA were significantly decreased. After wild-type or mutant hsa_circ_087631 vector and hsa-miR-346 mimics were transfected into 293T cells， the results of dual-luciferase reporter assay showed that hsa-miR-346 significantly decreased the luciferase activity of wild-type hsa_circ_087631 （P<0.01）， but the regulation did not change significantly after mutation of the predicted binding site. CONCLUSION Peripheral blood hsa_circ_087631 level is elevated in the PBC patients. The hsa_circ_087631/hsa-miR-346/Bcl-6 signaling may take effect in human T cells. Hsa-miR-346 significantly reduces the expression of hsa_circ_087631， but it may not be regulated by predicted binding sites.     

甘蓝型油菜耐低温生长生理生化响应机制

为提高甘蓝型油菜耐寒育种过程中的筛选效率，研究甘蓝型油菜耐低温机理，以8个不同抗寒性的甘蓝型油菜为材料，对低温条件下各材料的生物量、叶绿素、脯氨酸和相对电导率进行测定，发现在室外平均气温2.75℃时，抗性材料的生物量、叶绿素和脯氨酸的累积量都显著高于敏感材料，相对电导率没有显著差异；当平均气温7.52℃回升至12.39℃，抗性材料和敏感材料生物量的累积量无显著差异；在恒定低温10℃/4℃处理下抗性材料和敏感材料在处理前3周生物量均持续累积，但从第4周开始敏感材料新叶出现白斑，生物量减少，抗性材料老叶出现白斑，生物量维持不变。结果表明，耐低温材料在低温条件下叶绿素含量受到的影响较小，脯氨酸的积累量持续上升，具有较强的快速适应能力，在低温下具有显著的持续生长优势。     

Reduction in cadmium accumulation in japonica rice grains by CRISPR/Cas9-mediated editing of OsNRAMP5

Cadmium (Cd) intake is harmful to human health and Cd contamination in rice grains represents a severe threat to those consuming rice as a staple food. Knockout of Cd transporters is a promising strategy to reduce Cd accumulation in rice grains. OsNRAMP5 is the major transporter for Cd and manganese (Mn) uptake in rice. Nevertheless, it is uncertain whether knockout of OsNRAMP5 is applicable to produce low Cd rice without affecting plant growth and grain yield. In this study, we adopted CRISPR/Cas9-based gene editing technology to knock out OsNRAMP5 in two japonica varieties. We generated three independent transgene-free osnramp5 mutants and investigated the effect of osnramp5 mutations on Cd accumulation and plant growth. Hydroponic experiments showed that plant growth and chlorophyll content were significantly reduced in osnramp5 mutants at low Mn conditions, and this defective growth in the mutants could be fully rescued by supply of high levels of Mn. Cd and Mn accumulation in both roots and shoots was markedly reduced in the mutants compared to that in wild-type plants. In paddy field experiments, although Cd in flag leaves and grains was greatly reduced in osnramp5 mutants, some agronomic traits including plant height, seed setting rate, and grain number per panicle were affected in the mutants, which ultimately caused a mild reduction in grain yield. The reduced plant growth in the mutants can be attributed to a marked decrease in Mn accumulation. Our results reveal that the manipulation of OsNRAMP5 should be treated with caution: When assessing the applicability of osnramp5 mutants, soil pH and soil water content in paddy fields need to be taken into consideration, since they might affect the levels of available Mn in the soil and consequently determine the effect of the mutation on grain yield.     

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...     

应用Excel软件分析敌敌畏农药对两广二号家蚕的安全间隔期

应用Excel软件中的曲线方程分析判断敌敌畏农药对两广二号家蚕的安全间隔期。结果表明：敌敌畏1000倍液的安全间隔期是6d多，其数学模型为幂函数曲线方：y=88．0653x-1834本方法克服以往家蚕的农药安全期无数学模型可循或使用的BASIC方法陈旧繁杂的缺点，也避免英文统计软件SPSS难以在农村普及的不足，是一种计...     

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。