正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

New Polyketides from the Antarctic Fungus Pseudogymnoascus sp. HSX2#-11

The species Pseudogymnoascus is known as a psychrophilic pathogenic fungus with a ubiquitous distribution in Antarctica. Meanwhile, the study of its secondary metabolites is infrequent. Systematic research of the metabolites of the fungus Pseudogymnoascus sp. HSX2#-11, guided by the method of molecular networking, led to the isolation of one novel polyketide, pseudophenone A (1), along with six known analogs (2–7). The structure of the new compound was elucidated by extensive spectroscopic investigation and single-crystal X-ray diffraction. Pseudophenone A (1) is a dimer of diphenyl ketone and diphenyl ether, and there is only one analog of 1 to the best of our knowledge. Compounds 1 and 2 exhibited antibacterial activities against a panel of strains. This is the first time to use molecular networking to study the metabolic profiles of Antarctica fungi.     

Identification of an H1N1 subtype of swine influenza virus and serological analysis

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.     

鸡的微卫星DNA标记与胴体性状的相关分析

测定了青脚麻鸡与隐性白鸡杂交后自交的F2代个体的全净堂重、全净堂率、脂肪宽带、腹脂重和腹脂率6个胴体性状,检测10个微卫星DNA标记,对标记基因型胴体性状值进行方差分析和多重比较,结果表明:MCW328对于10周龄体重和全净膛重两性状各基因型均有显著的差异(P<0.05);ADL292对于腹脂重性状各基因型有极显著的差异(P<0.01),对于腹脂率有显著的差异(P<0.05)。MCW328标记基因与控制10周龄体重和全净膛重QTL连锁,ADL292标记基因与控制腹脂重和腹脂率QTL连锁。     

S1核酸酶突变检测法的优化

优化了S1核酸酶突变检测体系,结果表明(1)扩增法比杂交法产生异源双链DNA更节省时间;(2)PCR产物可以直接作为突变检测的底物;(3)适当降低反应温度、适当增加反应缓冲液中的NaCl浓度、适当缩短反应时间可提高突变检测效果。     

Genetic diversity of herbicide-resistant and -susceptible Avena fatua populations in North Dakota and Minnesota

Genetic diversity within and among 20 herbicide-resistant (HR) and 16 herbicide-susceptible (HS) Avena fatua multi-field populations was determined using 82 polymorphic loci resulting from two intersimple sequence repeat (ISSR) primers and one long-primer random amplified polymorphic DNA (LP-RAPD) primer. Collections from the Red River Valley of North Dakota and Minnesota, sampled in 1964 and 2000, represented A. fatua populations before and after intensive exposure to herbicides. A 1995 collection from south-west North Dakota represented A. fatua exposed to low herbicide selection. Despite differences in years of herbicide exposure among collections, both HR and HS populations from every collection maintained nearly similar levels of ISSR and RAPD diversity. Genetic differentiation among populations (G_ST) varied from 11% to 13% among HR populations and from 9% to 16% among HS populations, indicating that 84–91% of total variation remained within HS or within HR populations. Minimal difference in gene diversity between HR and HS is consistent with multiple origins of resistance, where HR A. fatua most likely evolved from diverse founding individuals.     

花生DNA导入大豆育种效果的研究

利用花粉管通道在大豆自花授粉后将花生ＤＮＡ导入栽培大豆受体中，引起受体的结荚习性、株高、成熟期，主茎节数、分枝数、产量性状和化学品质等性状的广泛变异。对变异株进行选择，获得了产量比受体提高１１．９％～２５．１％，蛋白质含量提高３．９％～５．３％的变异株系。实验结果表明：利用外源ＤＮＡ导入技术进行生态性状、产量性状和化学品质方向的育种是可能的。     

分子间的作用及其结构适应性

某些分子间的作用无法的用范德华力、氢键、介电常数、偶极矩等化学基本理论来加以解释，引入控制论的方法对这些分析间的作用进行研究后的结果表明，选择性分析间引力很可能是分子是最强的物理作用，而分子间的化学作用亦与分子间的结构适应性密切相关。     

分子标记用于赤眼蜂分子监测的研究

通过对松毛虫赤眼蜂（Trichogramma dedrolimi Matsumura）以及玉米螟赤眼蜂（Trichogramma ostriniae Pang et Chen）rDNA-ITS2基因的克隆测序，并联机检索GenBank核酸序列库中其它赤眼蜂的相关序列，利用核酸分析软件找到松毛虫赤眼蜂和玉米螟赤眼蜂ITS2序列中一段有鉴别意义的标志，并据此设计出蜂种的特异PCR引物，实现了松毛虫赤眼蜂和玉米螟赤眼蜂rDNA特异区带的PCR扩增。研究表明该特异区带具有种的特异性。我们认为该分子标记技术可用于目前我国生防中常用的松毛虫赤眼蜂和玉米螟蒌眼蜂的分子监测，如田间种群动态监控和寄生效果评估。