Investigation and genetic mapping of a Glomerella leaf spot resistance locus in apple

Apple Glomerella leaf spot (GLS) is a severe fungal disease that damages apple leaves during the summer in China. Breeding new apple varieties that are resistant to the disease is considered the best way of controlling GLS. Fine mapping and tightly linked marker are critically essential for the preselection of resistant seedlings. In this study, a population of 207 F₁ individuals derived from a cross between ‘Golden Delicious’ and ‘Fuji’ was used to construct a fine simple sequence repeat (SSR)‐based genetic linkage map. The position of R_gls, a locus responsible for resistance to GLS, was identified on apple linkage group (LG) 15 using SSR markers CH05g05 and CH01d08, which was adapted from a published set of 300 SSR markers that were developed using the bulked segregant analysis (BSA) method. These two SSR markers flanked the gene, and its recombination rate was 8.7% and 23.2%, respectively. A total of 276 newly developed SSR markers around the target region and designed from the genome apple assembly contig of LG15 were screened. Only nine of these were determined to be linked to the R_gls locus. Thus, a total of 11 SSR markers were in linkage with R_gls, and mapped at distances ranging from 0.5 to 33.8 cM. The closest marker to the R_gls locus was S0405127, which showed a genetic distance of approximately 0.5 cM. The first mapping of the gene R_gls was constructed, and the locations of the 11 effective primers in the ‘Golden Delicious’ apple genome sequence were anchored. This result facilitates better understanding of the molecular mechanisms underlying the trait of resistance to GLS and could be used in improving the breeding efficiency of GLS‐resistant apple varieties.     

Agronomic traits of soybean cultivars released in different decades after grafting record‐yield cultivar as rootstock

Breeders have seldom considered the selection for root traits during the genetic improvement in soybean. It is hypothesized that grain yield would be increased by the root function improvement, especially for the current cultivars. The objective of this grafting experiment was to determine the effect of record‐yield cultivars L14 or Z35 as rootstocks on agronomic traits of cultivars released in different decades. A total of 11 cultivars, released in different decades, were used to graft onto L14 or Z35 rootstocks. The agronomic traits were measured in the pot‐culture experiments. Grafting cultivars released in different decades onto L14 or Z35 rootstocks resulted in higher yield, 100‐seed mass and apparent harvest index as compared with those of non‐grafted or self‐grafted plants. Grain yield gain of cultivars grafted onto record‐yield cultivar rootstocks was 0.40 g/plant/year from 1966 to 2006, which was larger than that of non‐grafts and self‐grafts (0.27 g/plant/year). The yield of current cultivars should increase more if their root functions were improved.     

TRMM在海河流域南系的降水估算精度评价及其对SWAT模型的适用性

准确估算区域降水对水文过程评价和水资源管理意义重大。为评估TRMM 3B42V7降水产品在海河流域南系的估算精度及其在土壤和水评估模型(Soil and Water Assessment Tool,SWAT)中的适用性,利用28个气象站降水观测数据(2007-2016年)和101个雨量站观测数据(2010-2016年)开展研究。研究表明:站点尺度上,3B42V7降水产品对月降水估算的均方根误差小于15 mm,平均误差小于8.5 mm;在湿润季节的估算精度更好。流域尺度上,日降水估算精度较差,相关系数小于0.6。分区尺度上,3B42V7能够很好地捕捉到不同等级降水强度,但对微量降雨有所低估;山区和平原的年降水量均出现高估现象,平原区较为突出;此外,3B42V7能够较好地捕捉到研究区内极端降水的时间和空间分布。分2种情景进行水文模拟,利用月平均流量对模型进行校准和验证,在情景Ⅰ中,验证期模拟结果较好,决定系数在0.56~0.96之间,纳什效率系数在-11.09~0.94之间。TRMM 3B42V7可为海河流域及其类似区域的水资源管理提供参考。     

荷斯坦牛新遗传缺陷单倍型HH7分子筛查

HH7(Holstein Haplotype 7)是在荷斯坦牛群中新发现的一种隐性遗传缺陷单倍型,由27号染色体上CENPU基因4个碱基缺失突变引起,隐性基因纯合时能引起胚胎早期流产,严重影响着奶牛养殖者的经济效益.本研究利用竞争性等位基因特异性PCR(KASP法)对随机抽取的166头荷斯坦公牛样本进行了检测分析,结果...     

利用风洞实验研究~7Be示踪估算土壤风蚀速率的可行性

以黄土高原水蚀风蚀交错带沙黄土为研究对象,利用室内风洞模拟实验对7Be示踪估算土壤风蚀速率的可行性进行探讨。由于风蚀过程易带走土壤细颗粒,且7Be在土壤细颗粒中含量较高,所以利用7Be水蚀模型计算的土壤风蚀速率高于实测值。实验中发现样点风蚀后和风蚀前土壤颗粒比表面积之比与样点风蚀后7Be含量之间存在幂函数关系,基于此,提出颗粒校正系数(P)的计算式,并将P引入到7Be水蚀模型对其进行修正。计算分析发现,和实测值相比,利用修正模型计算的土壤风蚀速率误差均不超过5%,这说明经过修正的7Be水蚀模型能较准确地估算土壤风蚀速率,利用7Be示踪技术估算土壤风蚀速率是可行的。研究为进一步利用7Be示踪技术调查黄土高原水蚀风蚀交错带土壤风蚀问题奠定了基础。     

猪繁殖与呼吸综合征病毒ORF7基因的表达和重组蛋白的纯化及免疫学活性分析

[目的]在基因工程菌中实现猪繁殖与呼吸综合征病毒（PRRSV）ORF7基因的高效表达,对表达的重组蛋白进行纯化,并探讨其免疫学活性及应用价值。[方法]将已构建好的重组表达载体pET-ORF7转化大肠杆菌BL21（DE3）,用IPTG在最适条件下诱导表达,其表达产物用SDS-PAGE和Western Blot进行检测。应用Ni-NTAHis.Bind Resin层析柱在变性条件下纯化表达产物,通过梯度透析进行复性。采用Western Blot及间接ELISA法检测复性后重组蛋白的免疫学活性。[结果]重组质粒pET-ORF7在大肠杆菌中以融合形式成功表达,融合表达产物主要以包涵体形式存在。SDS-PAGE检测所表达的重组蛋白分子量为33 kD左右,与预期大小相符。Band-scan软件对表达蛋白分析发现,表达产物约占菌体蛋白的50%。Western Blot及间接ELISA法显示复性后的重组蛋白与PRRSV阳性血清有特异性结合反应,表明其具有良好的免疫学活性。[结论]该试验将为进一步研制PRRSV单克隆抗体及诊断试剂盒奠定基础。     

Cr6+、Mn7+和Hg2+对青虾的毒性和联合毒性研究

探讨了Cr6 、Mn7 和Hg2 三种重金属离子对青虾(Macrobrachium nipponense)的急性毒性及联合毒性作用。结果表明:Cr6 对青虾的24、48、72、96h的半致死浓度LC50分别为8.937、5.001、3.484、2.241mg/L;Mn7 对青虾24、48、96 h的LC50分别为45.497、34.191、13.293、2.460mg/L;Hg2 对青虾的24、48、72、96 h的LC50分别为0.0415、0.0239、0.0168、0.0131mg/L。急性毒性由强至弱依次为:Hg2 >Cr6 >Mn7 。Cr6 -Hg2 对青虾96 h的联合作用表现为协同作用;Mn7 -Hg2 对青虾的96 h联合急性毒性表现为:低浓度的Mn7 对Hg2 表现为协同作用,低浓度的Hg2 对Mn7 表现为拮抗作用;Mn7 -Cr6 96 h联合急性毒性表现为:高浓度的Mn7 对Cr6 表现为拮抗同作用,而非低浓度的Cr6 对Mn7 均表现为协同作用。并就Cr6 、Mn7 和Hg2 对青虾的急性致毒效应特征、安全浓度以及重金属离子间联合毒性效应进行了分析与探讨。     

巴西橡胶树热研7-33-97·PR107·RRIM600生理特性比较

[目的]为热研7-33-97推广提供科学依据。[方法]以3个橡胶树无性系热研7-33-97,PR107,RRIM600为试材,研究了不同品系的干胶含量,排胶初速度,堵塞指数,蔗糖,硫醇,无机磷,镁离子含量,黄色体破裂指数等生理指标。[结果]RRIM600、热研7-33-97和PR107的干胶总产量分别为538.725、26.89和409.37 g。3个品种干含依次为PR107>RRIM600>热研7-33-97。热研7-33-97蔗糖含量介于RRIM600和PR107之间。RRIM600硫醇含量较低,热研7-33-97与PR107硫醇含量较高。热研7-33-97与RRIM600无机磷含量较高。热研7-33-97镁离子含量最高。破裂指数以热研7-33-97最高,RRIM600最低。品系PR107的堵塞指数在各个阶段均明显高于热研7-33-97与RRIM600。[结论]热研7-33-97是一个较耐刺激且高产的品系。     

猪TAF7基因的克隆、染色体定位和组织表达谱分析

 【目的】通过对猪TAF7基因初步的研究,为猪分子遗传育种提供基础分子生物学信息,为猪的遗传育种提供分子标记。【方法】以五指山猪为研究对象,克隆TAF7基因,分析该基因结构特点,然后利用IMpRH（the INRA-University of Minnesota porcine radiation hybrid,法国农业科学院-明尼苏达大学的辐射杂种克隆板）分析该基因在猪染色体上定位信息,利用半定量RT-PCR方法,分析该基因在成年五指山猪16个不同组织（心脏、背肌、淋巴、脾脏、肝脏、肾脏、肺脏、子宫、睾丸、胃、小肠、大肠、卵巢、胸腺、脑、脂肪）的表达谱信息。【结果】克隆得到长1 701 bp的五指山猪TAF7基因序列,其中包括1 050 bp完整CDS（Coding Sequence,编码序列）区域,分析表明其编码含349个氨基酸的蛋白质。利用IMpRH分析结果表明,猪TAF7基因与分子标记SW1879和IL4（interleukin-4,白细胞介素-4）紧密连锁,LOD（Limit of Detection,检测极限）值分别为6.69和6.15。组织表达谱分析结果显示该基因在大多数组织中均有表达,其中在睾丸表达量较高,而在心脏、背肌中表达很低。【结论】猪TAF7基因5’UTR中有一个短的内含子,而在该基因CDS区域没有内含子。猪TAF7蛋白序列与人TAF7蛋白序列的相似性较高,二者在生物系统发育树中的距离最接近。猪TAF7基因在大多数组织中均表达,在猪睾丸中表达量较高,证实它调节目的基因转录具有广泛性。     

牛轮状病毒VP4基因与LTB基因的融合表达及免疫原性研究

PCR扩增轮状病毒VP4基因和大肠杆菌LTB基因编码区,将其克隆至pET32a载体上,构建pET32a-VP4-LTB质粒,转化大肠杆菌BL21(DE3),PCR、酶切鉴定后进一步测序,测序结果表明融合基因片段由2637 bp组成,为编码879个氨基酸残基的多肽。经IPTG诱导和SDS-PAGE检测发现表达产物分子量约为118.9 KD,以包涵体的形式存在。融合蛋白经镍离子螯合次氨基三乙酸(Ni-NTA)亲和层析纯化后免疫小鼠,结果所产生的抗体效价高于单独VP4蛋白的免疫结果但差异不显著(P0.5),但与对照组相比差异显著。表明所表达的融合蛋白具有较好的免疫原性,为研制牛轮状病毒亚单位疫苗奠定了基础。