大麦籽粒淀粉和β-葡聚糖积累特性研究

为研究大麦籽粒支链淀粉、直链淀粉和β-葡聚糖积累特性,及淀粉各组分与β-葡聚糖的关系,以2个皮大麦、1个裸大麦和1个糯裸大麦为试验材料,测定花后7、14、21、28d淀粉各组分及β-葡聚糖含量。结果表明,甘啤6号淀粉各组分含量随灌浆推进均逐渐升高,在成熟期达到最大值;甘垦啤7号和甘垦6号均呈先升高后降低趋势,在花后21d有1个峰值。糯大麦C 2-1直链淀粉含量显著低于非糯大麦;支链淀粉含量显著高于非糯大麦,整个灌浆期呈先升高后降低趋势。β-葡聚糖含量均随灌浆时间延长逐渐升高,成熟期含量最高;整个灌浆期糯大麦C 2-1含量显著高于非糯大麦。相关分析表明,直链淀粉/支链淀粉比值与β-葡聚糖含量呈极显著负相关,可以将直链淀粉/支链淀粉比值作为高β-葡聚糖品种选育的一个指标。Logistic方程拟合发现,直链淀粉、β-葡聚糖最终积累量与积累起势与有效积累时间有关,支链淀粉最终积累量取决于最高积累速率和平均积累速率。     

瓜类褪绿黄化病毒及其系统进化分析

【目的】检测新疆南疆7个县/市瓜类褪绿黄化病毒(Cucurbit chlorotic yellows virus, CCYV),进行系统发育分析,为新疆南疆CCYV预警和防控提供参考。【方法】于2019年,从中国新疆南疆7个县/市采集带有黄化特征的51份甜瓜叶片样品,RT-PCR进行CCYV的检测,进行特异性片段的克隆、测序和系统发育分析。【结果】中国新疆巴楚县、阿克苏市、莎车县、伽师县、疏勒县、疏附县均未检测到CCYV,而洛浦县的13份样本中,8份检出为CCYV阳性,检出率为61.53%,洛浦县CCYV特异性片段克隆测序获得了685 bp的核酸序列,与GenBank中的CCYV分离物外壳蛋白(Coat protein, CP)的一致性达到100%。所得序列与中国新疆(吐鲁番)、中国其他省、苏丹、日本、塞浦路斯、黎巴嫩的CP基因聚在I组(Group),沙特阿拉伯的分离物聚在Ⅱ组,伊朗的分离物聚在Ⅲ组。【结论】CCYV在中国新疆南疆洛浦县甜瓜产区发生,与中国新疆吐鲁番、中国其他省及周围国家的CCYV分离物亲缘关系很近,且群体遗传变异很小。     

Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

Genome wide association studies (GWAS) of element contents in grain with a special focus on zinc and iron in a world collection of barley (Hordeum vulgare L.)

Genome wide association studies (GWAS) were carried out to map Quantitative Trait Loci (QTL) associated with element contents in the grain using 336 spring barley. Of the elements analyzed, Fe content ranged from 21.9 to 91.0 mg kg⁻¹, Zn from 10.4 to 54.5 mg kg⁻¹, Ba from 0.2 to 8.9, Ca from 186.4 to 977.5, Cu from 1.5 to 9.8, K from 353.2 to 7721.5, Mg from 1049.8 to 2024.2, Mn from 8.1 to 22.9, Na from 55.9 to 627.9, P from 2272.9 to 5428.8, S from 880.7 to 1898.0, Si from 19.1 to 663.2, and Sr from 0.35 to 2.62 mg kg⁻¹. GWAS were carried out using 6519 SNP markers and multiple elements in MLM:PCA + K model in TASSEL software. Population analyses showed two sub-populations, primarily based on row types. GWAS for row types showed association with INTERMEDIUM-C, a modifier gene for lateral spikelet fertility in the 4H chromosome, validating current GWAS approach. GWAS also showed that 2 QTL for Ba, 2 for Ca, 4 for Cu, 11 for Fe, 2 for K, 3 for Mg, 6 for Mn, 4 for Na, 3 for S, 5 for Si, and 3 for Zn were mapped in barley chromosomes. The QTL identified in the current study are valuable for breeding nutrient dense barley cultivars in the future, especially Zn and Fe.     

Detect unique molecular structure associated with physiochemical properties in CDC varieties of oat grain with unique nutrient traits [Feed Type vs. Milling Type] in comparison with barley grain using advanced molecular spectroscopy as a non-destructive biological tool

The objective of this study was to determinate grain unique protein inherent molecular structure that are related physiochemical and nutrient profiles in CDC developed oat varieties [CDC Nasser (Feed Type) and CDC Seabiscuit (Milling Type)] grown in cool climate condition in western Canada in comparison with conventional barley variety of CDC Meredith as a control using advanced molecular spectroscopy. Multivariate analyses, including an agglomerative hierarchical cluster analysis (CLA) and principal component analysis (PCA), were performed to identify protein molecular structural differences among the grains. The results revealed that CDC Seabiscuit contained greater (P < 0.05) protein structural Amide I and II than CDC Nasser and CDC Meredith, while the greater (P < 0.005) structural Amide I to II area and height ratios was detected in CDC Meredith. New oat grains had greater (P < 0.05) β-sheet height than barley grains, however, there was no difference in α-helix to β-sheet ratio values among the varieties. In conclusion, CDC Nasser and CDC Meredith had no difference in protein molecular structural features, while CDC Seabiscuit contains different protein structural characteristics as compared to CDC Meredith grain. The molecular structure features are highly associated with physiochemical and nutrient profiles in grains, which indicate that it also affect nutrient utilization and availability.     

寨卡病毒SYBR GreenⅠreal-time PCR方法的建立及应用

为建立一种针对寨卡病毒的快速诊断方法,本研究根据寨卡病毒的3’端保守基因序列,设计合成1对引物,建立了检测寨卡病毒的荧光定量PCR方法。结果显示:所建立的检测方法的Ct值与标准品在1.41×10^1~1.41×10^10^ copies/μL具有良好的线性关系,相关性为1,斜率为-3.502;灵敏性结果显示,该方法的检测限度为1.41×10^1 copies/μL,是普通PCR的10000倍;特异性结果显示,对CHIKV、DENV和JEV无特异性扩增,特异性强;重复性试验结果显示,组内和组间变异系数均小于1%,重复性好。本研究建立的SYBR Green I real-time PCR检测方法,可用于寨卡病毒感染的快速诊断。     

Milk vetch dwarf virus infection in the Solanaceae and Caricaceae families in Southeast Asia

Milk vetch dwarf virus (MDV) is an important member of the genus Nanovirus and is transmitted by the aphid Aphis craccivora. MDV has multiple single-stranded DNA genome components, each approximately 1 kb, and two or three alpha-satellite molecules. It mainly infects plants of the legume family Fabaceae. Recently, papaya (Carica papaya) collected in Yesan, South Korea, displaying symptoms of leaf yellowing and dwarfism, was identified as a new host for MDV. To examine the geographical distribution of MDV, papaya samples with symptoms were harvested in South Korea, Vietnam, and Taiwan in August 2018, along with tomato and pepper samples grown in adjacent fields in Vietnam. The results revealed the presence of MDV not only in papaya but also in pepper and tomato. This MDV infection in members of the Solanaceae family was confirmed by Southern blot hybridization performed using a PCR product of segment S as a probe. Based on sequence analysis of three MDV components (M, S, and C3), we verified the presence of three different isolates of MDV in these three countries and homology between sequences of isolates from papaya and from members of the Solanaceae from Vietnam. Taken together, our results clearly demonstrate MDV infection in Vietnam and Taiwan for the first time and confirm that MDV can infect economically important pepper and tomato.     

利用酵母双杂交系统筛选与草莓镶脉病毒移动蛋白P1互作的寄主因子

 构建感染草莓镶脉病毒(SVBV)森林草莓的酵母cDNA文库,利用酵母双杂交系统,筛选出与SVBV P1蛋白互作的15种寄主因子。生物信息学分析发现,这15种寄主因子参与茉莉酸途径、泛素化、光合作用、抗病抗逆、蛋白修饰、蛋白运输和氧化还原等多种生物过程。另外,这些寄主因子还具有其他分子功能,包括氧化还原酶活性、蛋白二硫化物异构酶活性和金属离子结合活性等。本研究初步探讨了P1与寄主因子的互作机理,为揭示SVBV侵染森林草莓以及SVBV在寄主中扩展的分子机制提供理论依据。     

1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。     

一种基于RPA的番茄褪绿病毒检测方法

番茄褪绿病毒Tomato chlorosis virus(ToCV)引起番茄褪绿病毒病,给番茄生产造成严重危害。开发快速准确的检测方法对该病害的防控具有重要意义。利用番茄褪绿病毒外壳蛋白(CP)基因序列,设计特异性引物,建立了ToCV的重组酶聚合酶等温扩增(recombinase polymerase amplification, RPA)检测方法,同时分析了该方法的灵敏度和特异性。结果表明,建立的ToCV-RPA方法在38℃恒温下40 min可从ToCV阳性的番茄样品中扩增出246 bp的特异性条带。扩增时间短,对设备要求低,且与番茄其他病毒无交叉反应,特异性好,灵敏度可达到PCR方法的10倍,适用于ToCV的快速检测。