过表达ZmIBH1-1提高玉米苗期抗旱性

【目的】干旱是严重影响玉米生长发育进程的一个重要因素。挖掘玉米抗旱相关基因,通过转基因功能验证和转录组分析,解析关键基因在响应干旱胁迫过程中的分子调控机制,为抗旱分子育种和遗传改良提供理论依据。【方法】以玉米自交系B104（WT）为背景材料,利用农杆菌介导方法构建过表达ZmIBH1-1转基因株系（ZmIBH1-1-OE）;通过对转基因植株进行草铵膦抗性筛选、标记基因和目的基因PCR检测,以及运用实时荧光定量PCR检测目的基因的表达情况,鉴定阳性植株和株系;以WT和ZmIBH1-1-OE转基因株系为材料,通过干旱处理（20% PEG6000）,进行表型鉴定和耐旱生理生化指标测定,验证ZmIBH1-1的抗旱功能;通过对干旱胁迫下玉米4叶期转录组的比较分析,鉴定出差异表达的基因（differentially expressed genes,DEGs）;结合DAP-seq（DNA affinity purification sequencing）分析,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的下游靶基因,利用基因组可视化软件IGV（integrative genomics viewer）分析ZmIBH1-1蛋白结合候选靶基因的位置,然后通过Dual-Luciferase试验验证ZmIBH1-1蛋白与靶基因的调控关系。【结果】通过玉米遗传转化获得12个转化事件;T₃代中,能同时检测到标记基因Bar和目的基因ZmIBH1-1的植株有458个,实时荧光定量PCR检测结果表明,ZmIBH1-1-OE中ZmIBH1-1的表达量显著高于WT,株系3和株系8表达量最高,将其自交获得T₄代转基因株系用于后续试验。在干旱胁迫条件下,ZmIBH1-1-OE株系存活率、叶片相对含水量、叶绿素含量、可溶性蛋白含量及其生理生化指标（超氧化物歧化酶、过氧化物酶、过氧化氢酶活性）均显著高于WT,说明玉米中过量表达ZmIBH1-1赋予玉米更高的耐旱性。转录组分析结果表明,WT与ZmIBH1-1-OE株系在干旱胁迫下有1 214个差异表达基因;Gene Ontology（GO）功能富集分析结果表明,差异表达基因主要涉及生物过程、细胞组分和分子功能,如在生物过程中主要涉及到光合作用、应激响应、脱水响应等;KEGG富集分析表明,差异表达基因主要参与植物激素信号传导、新陈代谢等过程。结合转录组显著差异表达基因和DAP-Seq分析所得到ZmIBH1-1蛋白的靶基因,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的11个候选靶基因,包括2个钙信号相关基因、3个半胱氨酸代谢相关基因、1个bHLH转录因子、1个应激响应蛋白、1个谷胱甘肽转移酶、1个氧化还原过程蛋白和2个乙烯响应因子;基因组可视化结果显示ZmIBH1-1蛋白可以结合靶基因启动子区;随后通过Dual-Luciferase试验进一步表明,ZmIBH1-1蛋白可以直接作用于11个候选靶基因,其中,ZmIBH1-1蛋白可以促进ZmCa-M、ZmSYCO、ZmbHLH54、ZmGlu-r1、ZmCLPB3和ZmP450-99A2的表达,抑制ZmAGD12、ZmCYS、ZmCYSB、ZmERF-107和ZmEIN3的表达。此外,在干旱胁迫下NAC、WRKY、MYB等转录因子在ZmIBH1-1-OE和WT株系中也存在差异表达。【结论】ZmIBH1-1的过表达可以增强玉米苗期的耐旱性;ZmIBH1-1蛋白通过直接调控乙烯信号通路中的ZmERF-107和ZmEIN3的表达提高玉米的耐旱性;ZmIBH1-1蛋白通过直接调控钙信号相关基因ZmCa-M和ZmAGD12增强玉米的耐旱性;ZmIBH1-1蛋白可能通过间接调控NAC、WRKY、MYB等转录因子响应干旱胁迫。     

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.     

Association of TNF-α, IL-10 and IL-6 promoter polymorphisms in pulmonary tuberculosis patients and their household contacts of younger age group

Single nucleotide polymorphisms (SNPs) of cytokine genes have been found to be involved in the clinical outcome of Tuberculosis. The present study was aimed to identify the high risk genotypes in Tuberculosis patients and their household contacts. A total of 490 subjects were studied which includes 150 active pulmonary tuberculosis patients (APTB), 190 household contacts (HHC) and 150 healthy controls (HC). The SNPs of TNF-α (-308A/G), IL-10(-1082G/A) and IL-6(-174G/C) were performed by ARMs PCR. The IL-10 GA genotype showed significant association in APTB and HHC and was 2.3 times higher risk in APTB and 3.7 times in HHC compared to HCs. The A allele was found to be significantly associated with the risk of disease. The CC genotype of IL-6 was found to be significantly associated in APTB and an insignificant positive association in HHCs. The multifactor dimensionality reduction (MDR) analysis indicated that the genotypes of IL-6 were showing high risk with GA genotype of IL-10. In conclusion the gene interaction may be useful for identification of genotypes as biomarkers to distinguish high risk individuals.     

Effects of Benzyl Adenine and Abscisic Acid on Grain Yield and Yield Components in Triticale and Wheat

Grain yield and yield components (grains per ear, grain weight, 1000-grain weight, ear weight, ear seed ratio and dry matter partitioning between ear and seed) were examined in a wheat genotype (PBW-343) with well-filled grains and a Triticale genotype (DT-46) with poorly filled grains (showing grain shrivelling) grown in pots. Six days after anthesis (DAA), benzyladenine (BA) @2 µg ear⁻¹ and abscisic acid (ABA) @4 µg ear⁻¹ were injected at the base of the mother shoot ear in both species. It was observed that, in both wheat and Triticale , BA increased the grain weight, grain number and partitioning of dry matter between ear and seed, whereas ABA decreased the grain weight, grain number and dry matter partitioning between ear and seed. However, these decreases were slower in Triticale than in wheat. BA treatment increased the grain dry matter accumulation, which in turn resulted in better filling of grains and increased the grain weight in both wheat and Triticale . The average grain weight of Triticale was lower than that of wheat. Thus, it appears that variation in grain weight between wheat and Triticale might be due to different availabilities of growth-promoting phytohormones such as cytokinins and assimilates.     

辛亥革命前后张謇与袁世凯的交往

张謇与袁世凯的交往长达三十余年,他从最初的相识荐袁、拥袁合作、相互借重到后来的分离,张謇曾幻想依靠袁世凯实现自己“实业救国”的抱负,却以失败而告终。二人分分合合,充满波折的交往过程反映了中国近代资本主义发展的艰难与坎坷。     

试论工科院校大学生的审美教育

工科院校在强调专业教育的同时，没能同样重视审美教育。加强对工科院校大学生的审美教育，有助于培养正确的审美观，提高审美能力;有助于完善知识结构，提升人文精神; 有助于形成独立完善的人格，提升人格境界;有助于培养个性和谐、德智体美全而发展的高素质人才。     

BA—Dot—ELISA检测牛结核血清抗体的研究

利用BA—Dot—ELISA检测72份结核变反阳性牛血清,阳性率为69.5％,比常规ELISA(44.4％)高。用该法检测布鲁氏菌病、粘膜病、传染性鼻气管炎、白血病病牛血清及堪萨斯分枝杆菌、偶发分枝杆菌高免牛血清均无交叉反应,但对50份副结核变反阳性牛血清出现了一定的交叉。该法重复性好,简便快速,结果判定不需特殊仪器。     

夏谷新品种豫谷6号的选育

豫谷６号是应用有性杂交技术，经连续南繁、温室加代选择育成的夏谷新品种。具有高产稳产、高抗倒伏、高度抗旱、综合抗病力强等优良特性，适宜中等以上肥力水、旱地种植。     

流行性出血热病毒气溶胶感染乳鼠的免疫病理学研究

采用免疫组化ＢＡ法，对流行性出血热病毒（ＥＨＦＶ）气溶胶感染乳鼠脏器组织中的病毒动态分布进行了观察，并对其抗体消长进行了测定。结果表明：①ＢＡ法为病理诊断ＥＨＦＶ感染提供了一种准确的手段；②ＥＨＦＶ可通过气溶胶途径感染乳小鼠，乳小鼠感染ＥＨＦＶ的ＬＤ５０值可作为环境浓度的参考值；③ＥＨＦＶ气溶胶途径感染乳鼠后，４ｈ即可在心、肺，１ｄ在嗅球部检出，并可在其组织细胞中增殖，经血循、嗅神经、单核巨噬细胞系统等多途径扩散，造成多组织脏器的泛发性感染。感染后１２～１４ｄ，病毒检出率和病毒量均达峰值。动物自身免疫系统对ＥＨＦＶ有一定的清除能力；④感染乳鼠脑、肺、肾等脏器组织的病变与人类感染ＥＨＦＶ的病变有一定的相似性，提示可作为人ＥＨＦＶ的模型动物；⑤抗原检出率高于抗体检出率，ＥＬＩＳＡ法检出率高于ＩＦＡ法检出率。     

论散打侧踹腿的训练

侧踹腿是武术散打技术中最基本的腿法之一.速度快,力量大,不易防守,而且配合步法运用,变化多端,易于在不同距离上使用,既是进攻性腿法,也是防守性腿法.因此它在散打比赛中的作用非常重要.笔者结合多年的训练实践,阐述了侧踹腿的主要训练手段和在比赛中的合理运用.