Immunological Adjuvant Function of Aluminium Phosphate and Chicken IL-18 in NDV F Gene Vaccine

[Objective] This study aimed to investigate the immunological adjuvant function of aluminium phosphate and chicken IL-18 in NDV F gene vaccine. [Method] The vaccine （0.2 ml） containing aluminum phosphate adjuvant （90 μg）, pcDNA/F （200μg）, and pcDNA/chlL-18 （200 μg） was prepared. The 7 d old chick- ens to be tested were randomly divided into six groups （12 chickens in each group） and immunized through intramuscular injection with inactivated Newcastle disease vaccines, pcDNA/F＋pcDNA/chlL-18＋phosphate aluminum, pcDNA/F, pcDNA/F.＋pcDNA/ chlL-18, pcDNA/F＋aluminum phosphate, and physiological saline respectively; the secondary immunization was conducted with the same dose when the chickens were 21 d old. Their blood was sampled 0, 7, 14, 21, 28 d after first immunization. Anti- body titer was detected with ELISA and T cell transformation rate was measured with MIT. Experimental chicken will be challenged with 30 LD50 NDV virulence 28 d after first immunization. [Result] The survival rate of the chickens immunized with pcDNA/F＋aluminium phosphate＋pcDNA/chlL-18 achieved 8/12, higher than that of those immunized with pcDNA/F 4/12 and pcDNA/F＋pcDNA/chlL-18 （6/12）. The NDV antibody titer of the chickens immunized with pcDNA/F＋ aluminum phosphate, pcD- NA/F＋pcDNA/chlL-18 and pcDNA/F＋pcDNA/chlL-18＋aluminum phosphate is not differ- ent （P〉0.05）, but significantly lower than that of the chickens immunized with tradi- tional vaccine （P〈0.05）. The T cell transformation rate of the chickens immunized with pcDNA/F＋pcDNA/chlL-18＋aluminium phosphate was obviously higher than that of the chickens immunized with pcDNA/F （P〈0.05）. The T cell transformation rates of chickens immunized with pcDNA/F and the traditional vaccine showed no signifi- cant difference （P〉0.05）. [Conclusion] Combination of aluminium phosphate and pcD- NA/chlL-18 can significantly enhance the immune effect of NDV F gene vaccine.

The SNPs C.513A>T in the MHC B-F gene and rs15001532 in the SPOCK1 gene are associated with Salmonella pullorum disease resistance in chickens

supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China （CARS-41）;the National High Technology Research and Development Program of China （2011AA100301）

莲草直胸跳甲短神经肽F受体基因sNPFR的克隆及表达分析

为明确莲草直胸跳甲(Agasicles hygrophila)短神经肽F受体AhsNPFR功能及其表达特点,为探索莲草直胸跳甲生防作用奠定理论基础,利用PCR技术克隆鉴定莲草直胸跳甲AhsNPFR基因并进行生物信息学分析,通过实时荧光定量PCR技术分析其在莲草直胸跳甲不同发育时期和组织中的时空表达谱。结果表明,克隆获得基因AhsNPFR全长1 669 bp,开放阅读框1 257 bp,编码418个氨基酸;预测其蛋白质分子质量为48.11 ku,理论等电点为8.21,AhsNPFR具有7个典型保守跨膜结构域,属于GPCRs家族,系统进化树分析表明,其与玉米根萤叶甲Diabrotica virgifera virgifera sNPFR亲缘关系最近。AhsNPFR基因在不同发育阶段均有表达,在1龄幼虫中的表达量最高,是卵中表达量的9.06倍;在卵中表达量最低;雌成虫的表达量显著高于雄成虫。AhsNPFR基因在不同组织中均有表达,在3龄幼虫后肠中显著高表达,是脂肪体表达量的12.21倍;在雌雄成虫后肠显著高表达,并在所有组织中均没有雌雄表达差异。

蜡梅花发育相关基因CpUFO表达特性与功能分析

【目的】UFO是被子植物中首个被发现的F-box蛋白,主要参与调控花分生组织特性和花器官发育。本文通过对蜡梅CpUFO基因表达特性分析及异源表达拟南芥表型分析,初步鉴定CpUFO的功能,为阐明蜡梅花发育分子调控机制提供参考。【方法】基于前期构建的蜡梅转录组数据,克隆蜡梅CpUFO基因并对其编码蛋白进行同源比对及进化树分析。根据qRT-PCR分析CpUFO基因在不同组织、器官及全年花发育各阶段中的相对表达量,比较35S::CpUFO转基因拟南芥株系及Col-0野生型表型差异,并通过qRT-PCR检测过表达拟南芥株系内源开花相关基因的表达特性,分析过表达CpUFO对拟南芥开花时间以及花器官发育的影响。【结果】获得的CpUFO基因cDNA序列为1665 bp,编码403个氨基酸,含有保守的F-box结构域。CpUFO在外花被片中表达量相对最高,其次是内花被片,在果、茎、叶、腋芽及雌雄蕊中表达水平较低;而在全年花芽中的表达量分析结果显示,低温打破休眠后的露瓣和始花以及盛开期花芽中的表达量显著高于其他时期。与野生型拟南芥相比,35S::CpUFO转基因株系莲座叶数目减少,开花提前,且拟南芥内源基因TFL1表达量下调,成花关键基因FUL和AP1的表达量上调。【结论】CpUFO在不同组织、器官及花发育各阶段均有表达,不同组织中在外花被片中表达量最高,全年花芽中打破休眠的花蕾开放阶段表达量最高。同时,过表达CpUFO会促进拟南芥早花并影响其花器官发育。

6株鸽源新城疫病毒广东分离株F、HN基因的克隆及遗传进化分析

为研究广东省鸽源新城疫病毒(NDV)的遗传进化规律,试验于2021—2022年从广东6家疑似感染NDV的鸽场采集病鸽临床样品,进行RT-PCR诊断、病毒分离鉴定和动物回归试验,并对分离毒株进行生物学毒力测定以及F基因、HN基因的同源性和遗传进化分析。结果显示:共分离到6株鸽源NDV,对幼鸽进行分组攻毒14 d后,发病率均为100%,死亡率为60%~80%;6株毒株的F蛋白裂解位点氨基酸序列为112R-R-Q-K-R-F117/112R-R-R-K-R-F117,具有NDV强毒株的典型分子特征,和分离毒株ICPI的测定结果共同表明6株毒株均为强毒株;6株毒株均为NDV的ClassⅡ基因Ⅵ.2.1.1.2.2型;与传统疫苗株La Sota的同源性较低,且F、HN蛋白氨基酸出现多处位点变异。研究表明,广东省鸽源NDV流行以Class Ⅱ基因Ⅵ.2.1.1.2.2型为主,常用的疫苗株La Sota可能对广东地区流行的鸽源NDV保护效果不佳。

^(18)F-FDG PET/CT代谢参数联合临床病理特征对晚期结直肠癌患者KRAS基因突变的评估价值

目的探讨^(18)F-氟代脱氧葡萄糖(^(18)F-FDG)PET/CT代谢参数联合临床病理特征对晚期结直肠癌患者鼠类肉瘤病毒癌基因(KRAS)基因突变的评估价值。方法回顾性分析64例晚期结直肠癌患者的临床资料。所有患者在治疗前均接受^(18)F-FDG PET/CT检查。分析最大标准化摄取值(SUV_(max))、不同阈值下的代谢肿瘤体积(MTV)和病变总糖酵解(TLG)、临床病理特征与患者KRAS基因突变状态之间的关系。通过多因素Logistic回归模型分析与患者KRAS基因突变相关的因素。采用受试者工作特征曲线分析^(18)F-FDG PET/CT代谢参数、临床病理特征及二者联合评估患者KRAS基因突变的效能。基于^(18)F-FDG PET/CT代谢参数和临床病理特征构建列线图模型。结果单因素和多因素Logistic回归分析结果显示,SUV_(max)≥19.55、MTV50%≥7.95、肿瘤组织中/高分化与患者KRAS基因突变有关。SUV_(max)、MTV50%和肿瘤组织分化程度评估患者KRAS基因突变的曲线下面积(AUC)分别为0.653、0.625和0.621,三者联合评估的AUC为0.800,均高于单个指标的AUC(P<0.05)。基于SUV_(max)、MTV50%和肿瘤组织分化程度构建的列线图模型的一致性指数为0.800,校准曲线与参考线基本拟合。结论^(18)F-FDG PET/CT的代谢参数SUV_(max)、MTV50%与晚期结直肠癌患者KRAS基因突变密切相关,联合肿瘤组织分化程度所构建的列线图模型对患者KRAS基因突变具有较高的评估价值。

一体化^(18)F-FET PET/MR术前评估成人胶质瘤患者MGMT基因启动子甲基化状态

目的 旨在研究一体化^(18)F-氟乙基酪氨酸(^(18)F-fluoroethyltyrosine, ^(18)F-FET)正电子发射断层成像(positron emission tomography, PET)/MR对成人胶质瘤的O6-甲基鸟嘌呤DNA甲基转移酶(O6-methylguanine DNA methyltransferase, MGMT)基因启动子甲基化状态的鉴别能力。材料与方法 回顾性分析未经活检或治疗的胶质瘤患者资料16例,均完成一体化PET/MR扫描,包括^(18)F-FET PET、常规MRI及体素内不相干运动(intravoxel incoherent motion, IVIM)成像。以靶本比(target-background ratio, TBR)=1.6为阈值对PET图像进行感兴趣体积(volume of interest, VOI)分割,通过刚性配准获得肿瘤VOI对应的IVIM图及其参数表观扩散系数(apparent diffusion coefficient, ADC)、真实扩散系数(true diffusion coefficient, D)、伪扩散系数(pseudo diffusion coefficient, D^(*))、灌注分数(perfusion fraction, f)、分布扩散系数(distributed diffusion coefficient, DDC)和异质性指数(heterogeneity index, α),使用pyradiomics对各参数进行特征提取,获得对应的一阶灰度直方图特征。计算每个IVIM参数图对应的特征值与^(18)F-FET PET参数的关系,并利用组间比较和受试者工作特征(receive operating characteristic, ROC)曲线分析探究PET和IVIM参数对MGMT启动子甲基化的区分能力。结果 IVIM-α的两个特征值90分位值(r=0.526,P<0.05)和最大值(r=0.520, P<0.05)与PET参数平均标准摄取值(meanstandarduptakevalue,SUVmean)呈正相关;IVIM-α和SUVmean在MGMT启动子甲基化状态阳性和阴性两组之间的差异有统计学意义(P<0.05),阳性组的IVIM-α均值和SUVmean显著高于阴性组;融合IVIM-α均值和SUVmean对MGMT启动子甲基化状态进行区分的曲线下面积(area under the curve, AUC)为0.77。结论 基于一体化PET/MR的^(18)F-FET PET和IVIM参数能够有效预测胶质瘤的MGMT启动子甲基化状态。

Cloning and Sequence Analysis of HN and F Protein Genes from a Strain of Goose Paramyxovirus

[ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two pairs of pdmers were designed to amplify the HN and F genes from GX1 strain of goose paramyxovirus isolated from diseased goose in Guangxi Zhuang Autonomous Region; the amplified products were ligated into pMD18-T vector and sequenced. [ Result ] HN and F genes of this strain tested were 1 716 and 1 662 bp in full nucleotide length, respectively; both showed the homologues of about 97.3% with GPV- SF02 strain, of 80.3% -97.5% with strains LaSota, F48E9 and JS, of just 84.8% with Miyadera strain. [ Conclusion] The results show that isolated strain BX1 matches to virulent APMV-1 strain, belonging to genotype Ⅶ of APMV-1 strain.

兔源F型多杀性巴氏杆菌荧光定量PCR检测方法的建立

为建立特异性强且敏感性高的兔源F型多杀性巴氏杆菌(Pm)快速检测方法,本实验根据F型Pm fcbD基因的保守序列设计特异性引物和探针,经反应条件优化,建立了检测兔源F型Pm的荧光定量PCR方法。利用该方法检测兔源F型Pm、兔源A和D型Pm、支气管败血波氏杆菌、肺炎克雷伯菌、大肠杆菌、魏氏梭菌和金黄色葡萄球菌等临床常见兔源病原菌,结果显示,该方法仅对兔源F型Pm检测为阳性,其余病原菌均为阴性,特异性强。利用该方法分别检测10倍倍比稀释(1×10^(8)拷贝/μL~1×10^(0)拷贝/μL)的兔源F型Pm基因组DNA,进行敏感性试验,结果显示该方法的检测限为10拷贝/μL,敏感性分别是已报导的双重PCR方法和环介导等温扩增方法的100倍和10倍,敏感性高。利用该方法对不同稀释度的质粒标准品进行批内和批间重复性试验,结果显示,批内和批间变异系数均小于3%,重复性好。利用该方法检测64份已知结果的临床样品,结果显示该方法的检测结果与已报导的双重PCR方法和环介导等温扩增方法检测结果的符合率均为100%,准确性高。本研究首次以fcbD为靶基因建立兔源F型Pm的荧光定量PCR检测方法,为兔源F型Pm的检测提供了有力的技术手段。

大白菜F-BOX基因家族的鉴定与核盘菌诱导应答分析

【目的】筛选参与菌核病应答反应的F-BOX基因,为大白菜抗菌核病基因的功能研究及抗病品种的选育提供基础。【方法】基于核盘菌侵染大白菜转录组测序数据,对大白菜F-BOX基因家族进行鉴定,进行亚细胞定位、染色体定位、保守结构域等生物信息学分析。分析F-BOX基因在核盘菌侵染下的差异表达,并采用q-PCR技术检测F-BOX基因在0 h和36 h的表达情况。【结果】共鉴定32个BraF-BOX基因,分子量介于34 751.13~105 942.22Da;亚细胞定位表明,26个F-BOX基因定位在细胞核和细胞质中,6个定位在叶绿体中;系统进化树分析将BraFBOX分为4个亚族;基因结构分析结果表明,每个BraF-BOX家族成员均含有Motif 1,序列中均含有外显子;组织特异性表达分析结果表明在不同大白菜部位,表达量有差异;在核盘菌处理36 h时,q-PCR检测6个基因的相对表达量趋势与转录组数据一致,其中Bra037120、Bra011427、Bra009835等3个基因表达上调,核盘菌侵染时间越长,表达量越大。【结论】综合BraF-BOX基因生物信息学及转录组数据分析,Bra037120、Bra011427、Bra009835基因可能参与大白菜菌核病抗性功能,为后续基因功能研究提供研究基础。

基于F基因的鸽新城疫病毒分子特征分析

试验旨在了解鸽新城疫病毒流行株的分子特征,利用MEGA7.0、Lasergene7.0软件对2020—2022年新疆和田地区规模化养鸽场流行病学调查阳性样品进行了鸽新城疫病毒F基因的测序和序列分析。结果显示,6株F基因属于Ⅵ.2.1.1.2.2基因亚型,F蛋白裂解位点序列均为112R-R-Q-K-R-F117,与基因Ⅱ型疫苗株La Sota株、HB1株、VG/GA株、Clone30株及基因Ⅲ型Mukteswarz株的核苷酸同源性为83.5%~85.7%,与甘肃、宁夏鸽新城疫核苷酸同源性为97.3%~99.5%。6株F蛋白融合肽区均有两处氨基酸突变(V121I、A132S),HRa区均有1处氨基酸突变(V179I),NDV/PIGEON/XJHT/802/2022/F在HRb、HRc和跨膜区还有1~2个氨基酸突变。研究表明,该地区鸽新城疫病毒遗传相对稳定,变异持续存在,定期监测与分析鸽新城疫病毒分子流行特征十分必要。

怀化地区犬瘟热病毒H、F基因遗传多样性及其遗传进化分析

【目的】探究怀化地区犬瘟热病毒的基因变异与遗传多样性情况,阐明其遗传进化关系。【方法】从怀化地区收集30株犬源犬瘟热病毒样本,用PCR法扩增其H、F基因序列。利用DNAStar软件分析H、F基因序列碱基组成;利用Clustal X、MegAlign软件进行变异位点和遗传多样性分析;运用Clustal X、PhyML 3.0软件,采用最大似然法(ML)对怀化地区犬瘟热病毒进行遗传进化分析,并用FigTree v 1.3.1软件构建遗传进化树。【结果】30株怀化地区犬瘟热病毒均为Asia-Ⅰ型,其H、F基因序列长度分别为1778和1850 bp,其AT含量分别为56.8%~57.9%和54.9%~56.1%,GC含量分别为42.2%~43.0%和44.1%~44.9%。H基因的种内差异为0~3.1%,种间差异为51.36%~60.15%;F基因的种内差异为0~2.4%,种间差异为39.60%~53.09%。基于H、F基因序列所构建的遗传进化树发现,来源于怀化地区的30株犬源犬瘟热病毒全部位于遗传进化树的同一分支上。【结论】来源于怀化地区犬瘟热病毒之间虽存在一定程度的基因变异和遗传多样性,但它们之间的亲缘关系较近。且其H、F基因序列的种内差异小、种间差异大,是研究犬瘟热病毒基因变异、遗传多样性和遗传进化的重要遗传标记。

Relations of Budd-Chiari syndrome to prothrombin gene mutation

BACKGROUND: Budd-Chiari syndrome (BCS) is a type of disease characterized by portal hypertension and/or hy- pertension of the inferior vena cava (IVC) due to the ob- struction of the hepatic veins (HV) and/or intrahepatic IVC outlet. Being etiologically complicated and obscure, BCS can be acquired or idiopathic and several gene muta- tions may be contributable. This study was to explore whether prothrombin gene mutation (F G20210A) takes part in the pathogenesis of BCS and to investigate their cor- relativity. METHODS: In 38 proven BCS patients and 70 controls, polymerase chain reaction-restriction fragment length poly- morphism (PCR-RFLP) was used to find F G20210A mutation. To detect whether there are any mutations, four steps were taken: purification of genome DNA from whole blood, amplification of special fragment by polymerase chain reaction, digestion of the fragment via restriction en- donuclease, and analysis of results by polyacrylamide gel electrophoresis. RESULTS: F G20210A mutation was not detected in all patients and controls. CONCLUSIONS: No F G20210A mutation exists in Chi- nese patients with BCS, nor correlativity between the oc- currence of BCS and F G20210A mutation. The etiology of BCS in the Chinese needs further investigation.

Genetic Analysis and Molecular Mapping of a Stripe Rust Resistance Gene YrH9014 in Wheat Line H9014-14-4-6-1

Stripe rust, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most widespread and destructive wheat diseases in many wheat-growing regions of the world. The winter wheat translocation line H9014-14-4-6-1 has all stage resistance. To identify stripe rust resistance genes, the segregating populations were developed from the cross between H9014-14-4-6-1 and Mingxian 169 （a wheat cultivar susceptible to all Pst races identified in China）. The seedlings of the parents and F1 plants, Fz, F3 and BC1 generations were tested with Pst races under controlled greenhouse conditions. Two genes for resistance to stripe rust were identified, one dominant gene conferred resistance to SUN11-4, temporarily designated YrH9014 and the other recessive gene conferred resistance to CYR33. The bulked segregant analysis and simple sequence repeat （SSR） markers were used to identify polymorphic markers associated with YrH9014. Seven polymorphic SSR markers were used to genotype the F2 population inoculated with SUN11-4. A linkage map was constructed according to the genotypes of seven SSR markers and resistance gene. The molecular map spanned 24.3 cM, and the genetic distance of the two closest markers Xbarc13 and Xbarc55 to gene locus was 1.4 and 3.6 cM, respectively. Based on the position of SSR marker, the resistance gene YrH9014 was located on chromosome arm 2BS. Amplification of a set of nulli-tetrasomic Chinese Spring lines with SSR marker Xbarc13 indicated that YrH9014 was located on chromosome 2B. Based on chromosomal location, the reaction patterns and pedigree analysis, YrH9014 should be a novel resistance gene to stripe rust. This new gene and flanking markers got from this study should be useful for marker-assisted selection （MAS） in breeding programs for stripe rust.

禽4型副黏病毒F基因的克隆及原核表达载体构建

扩增禽4型副黏病毒(Avian paramyxovirus type 4,APMV-4)新疆野鸭源分离株APMV-4/Pintail/CH(XJ)/01/2016的F基因,并构建原核表达载体。参考GenBank上公布的APMV-4序列,通过DNAStar设计F基因特异性引物,以APMV-4/Pintail/CH(XJ)/01/2016基因组RNA为模板,采用RT-PCR方法扩增F基因编码区,连接于pMD19-T载体,用BamHⅠ和HindⅢ酶切原核表达载体pET-30a重组质粒,将酶切产物克隆至pET-30a多克隆位点上,转化至大肠杆菌DH5α感受态细胞中,构建F基因原核表达载体,并对阳性重组质粒进行PCR和测序鉴定。结果显示,扩增出的片段大小为1701 bp,经测序鉴定该扩增片段为F基因全长,与GenBank中收录的APMV-4序列核苷酸相似性为99%;连接表达载体结果表明,原核表达载体pET-30a-F成功构建,为APMV-4诊断方法的建立提供了良好的技术基础。

2020—2022年华中地区犬副流感病毒检测及其F、HN基因系统发育分析

为探究华中地区犬副流感病毒(CPIV)的流行及遗传进化情况,利用RT-PCR方法对华中地区2020年9月—2022年1月收集的418份病料样品进行了CPIV检测,并利用生物信息学软件对分离的CPIV毒株的F、HN基因进行同源性、氨基酸位点分析及系统发育分析。结果显示:CPIV阳性率为15.8%,扩增获得9株CPIV的F、HN基因的序列,并成功分离培养9株CPIV毒株;同时,将分离株的F、HN基因与19株副流感病毒参考毒株进行序列比对分析,结果表明,F基因核苷酸同源性为95.1%~100%,氨基酸同源性为94.6%~99.8%,HN基因核苷酸同源性为95.9%~99.9%,氨基酸同源性为95.8%~99.6%;CPIV的F及HN基因与参考毒株相比均有部分氨基酸发生了突变;系统发育分析结果显示,本研究获得的8株分离株与美国株CPI-和CPI+处于同一分支上,亲缘关系较近,另一株分离株CS-324与中国株He N0718和韩国株D277位于同一分支上。本研究丰富了华中地区CPIV的流行病学调查资料,并为该病的综合防控提供了理论依据。

Construction of Recombinant Expression Plasmids Containing H and F Protein Genes of Canine Distemper Virus Isolated from a Mink and Their Expression in Prokaryotic Cells

[Objective] This study aimed to construct the recombinant expression plasmids containing H and F protein genes of Canine distemper virus isolated from a mink and to express these two genes in prekaryotic cells as well as to study the reactogenieity of the expressed products. [ Method ] RT-PCR amplification was used to obtain H and F protein genes; TA cloning and subclonlng techniques were used to construct the cloning plasmids（pMD-18T-H and pMD-18T-F） and recombinant expression plasmids（pET28a-H and pET28a-F） ; SDS-PAGE and Western-blotting were adopted to verify whether the target proteins were successfully expressed. [ Result] The recombinant expression plasmids pET28a-H and pET28a-F containing H and F protein genes of Canine distemper virus isolated from a mink were successfully constructed, and both the expressed H and F proteins with respectively relative molecular mass of 31 400 and 38 200 produced positive reac- tion with the CDV standard positive serum. [ Conclusion] The H and F proteins expressed in prokaryotic cells were the same with the natural ones in terms of reac- togenicity, which can be utilized for diagnosis of a CDV＇s infection or for an epidemiological investigation. Meanwhile, they also provide a basis for developing ge- netically engineered subunit vaccines.

Inheritance and Molecular Mapping of Stripe Rust Resistance Gene Yr88375 in Chinese Wheat Line Zhongliang 88375

Stripe rust is one of the most important diseases of wheat worldwide. Inheritance of stripe rust resistance and mapping of resistance gene with simple sequence repeat （SSR） markers are studied to formulate efficient strategies for breeding cultivars resistant to stripe rust. Zhongliang 88375, a common wheat line, is highly resistant to all three rusts of wheat in China. The gene conferring rust disease was deduced originating from Elytrigia intermedium. Genetic analysis of Zhongliang 88375 indicated that the resistance to PST race CYR31 was controlled by a single dominant gene, temporarily designated as Yr88375. To molecular map Yr88375, a F2 segregating population consisting of 163 individuals was constructed on the basis of the hybridization between Zhongliang 88375 and a susceptible wheat line Mingxian 169; 320 SSR primer pairs were used for analyzing the genetic linkage relation. Six SSR markers, Xgwm335, Xwmc289, Xwmc810, Xgdmll6, Xbarc59, and Xwmc783, are linked to Yr88375 as they were all located on chromosome 5BL Yr88375 was also located on that chromosome arm, closely linked to Xgdmll6 and Xwmc810 with genetic distances of 3.1 and 3.9 cM, respectively. The furthest marker Xwmc783 was 13.5 cM to Yr88375. Hence, pedigree analysis of Zhongliang 88375 combined with SSR markers supports the conclusion that the highly resistance gene Yr88375 derived from Elytrigia intermedium is a novel gene for resistance to stripe rust in wheat. It could play an important role in wheat breeding programs for stripe rust resistance.

黄瓜F-box基因家族的鉴定与生物信息学分析

F-box蛋白是泛素连接酶E3的核心成分,在植物的生长发育和抵御胁迫方面起着重要作用。为探讨黄瓜F-box基因家族的分子特征与表达模式,通过同源序列比对,共鉴定出22个黄瓜F-box基因家族成员,在7条染色体上均有分布,依次命名为CsF-box01~CsF-box22。然后对该家族基因结构、染色体定位、理化性质、保守基序、顺式作用元件、进化关系和表达模式进行分析。结果表明,黄瓜F-box基因家族编码蛋白大多为不稳定亲水蛋白,55%的成员为酸性蛋白;保守基序分析发现,95%的成员含有Motif1,Motif1在该蛋白家族中最为保守;基因结构分析发现,该家族有8个基因为断裂基因,外显子数目在2~10个;亚细胞定位显示,F-box蛋白主要被定位到细胞质中,其次是细胞核中;启动子顺式作用元件分析发现,黄瓜F-box基因家族富含大量胁迫响应、生长发育调控、激素响应和光响应等作用元件,说明F-box基因可能广泛参与黄瓜生长发育和胁迫响应过程。基因表达分析结果发现,分别有8、2个F-box基因在霜霉菌和丁香假单胞菌胁迫下差异性表达,说明黄瓜F-box家族基因参与了生物胁迫响应途径。

Molecular Mapping of a Stripe Rust Resistance Gene YrH9020a Transferred from Psathyrostachys huashanica Keng on Wheat Chromosome 6D

Stripe rust （yellow rust）, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most devastating diseases of wheat throughout the world. H9020-1-6-8-3 is a translocation line originally developed from interspeciifc hybridization between wheat line 7182 and Psathyrostachys huashanica Keng and is resistant to most Pst races in China. To identify the resistance gene（s） in the translocation line, H9020-1-6-8-3 was crossed with susceptible cultivar Mingxian 169, and seedlings of the parents, F1, F2, F3, and BC1 generations were tested with prevalent Chinese Pst race CYR32 under controlled greenhouse conditions. The results indicated that there is a single dominant gene, temporarily designated as YrH9020a, conferring resistance to CYR32. The resistance gene was mapped by the F2 population from Mingxian 169/H9020-1-6-8-3. It was linked to six microsatellite markers, including Xbarc196, Xbarc202, Xbarc96, Xgpw4372, Xbarc21, and Xgdm141, lfanked by Xbarc96 and Xbarc202 with at 4.5 and 8.3 cM, respectively. Based on the chromosomal locations of these markers and the test of Chinese Spring （CS） nullitetrasomic and ditelosomic lines, the gene was assigned to chromosome 6D. According to the origin and the chromosomal location, YrH9020a might be a new resistance gene to stripe rust. The lfanking markers linked to YrH9020a could be useful for marker-assisted selection in breeding programs.