水稻OsUAP2基因生物信息学分析及原核表达活性鉴定

UAGPase广泛存在于生物体,是糖代谢过程中一类重要酶。我们前期发现水稻Os UAP1具有单向催化UDPG分解代谢活性,Os UAP2与Os UAP1序列相似性极高。本研究对Os UAP2进行生物信息学分析,构建并表达重组蛋白,并鉴定其酶蛋白活性。结果表明,OsUAP2基因位于4号染色体,编码区序列长1482 bp,编码493个氨基酸;Os UAP2具有UDPGP保守结构域,无跨膜区域,N-端不含信号肽;二级结构分析显示,该蛋白含α-螺旋和无规则卷曲较多;同源性分析表明,Os UAP2与玉米UAGPase的亲缘关系最近。在16℃下经0.1 mmol/L IPTG诱导8 h,原核菌株BL21表达出分子量约55 kD可溶性重组蛋白。体外酶活性测定表明,Os UAP2能同时催化UDPG降解和合成,表现出双向可逆催化的特点。本研究结果为进一步深入研究OsUAP2基因的生物学功能提供了科学依据。     

Heterosis and combining ability of seven maize germplasm populations

Understanding the combining ability and heterosis of available germplasm is a prerequisite for successful maize improvement and breeding. The objectives of this study were to analyze the combining ability and heterosis of seven representative maize germplasm populations, and further, to evaluate their potential utility in germplasm improvement. A total of 21 crosses were made among these seven populations in a complete diallel without reciprocals. The parental populations and 21 crosses were evaluated for days to silking (DS), ear height (EH), and grain yield (GY) in the Northeast and Yellow and Huai River maize growing areas in China in 2012. Csyn5, Csyn7, Cpop.11, and Cpop.12 had desirable general combining ability (GCA) effects for DS and EH in both the Northeast China mega-environment (NCM) and the Yellow and Huai River Regions of China mega-environment (YHCM). Cpop.11 possessed a favorable GCA effect for GY in the NCM, as did Csyn5, Cpop.17, and Cpop.18 in the YHCM. Csyn6 and Csyn7 exhibited tremendous yield-enhancing potential in both mega-environments. Additionally, six combinations including Csyn7 × Csyn6, Csyn5 × Csyn6, Cpop.11 × Cpop.18, Cpop.12 × Cpop.17, Csyn7 × Cpop.17, and Csyn5 × Csyn7 exhibited better specific combining ability effects for GY, yield performance, and mid-parent heterosis in the appropriate mega-environment. These results indicated that the seven populations would be very useful for the improvement of related agronomic traits, and the six candidate combinations possessed great potential for further improvement and utilization.     

中子辐照对锆合金显微组织的影响研究进展

作为反应堆的第一道安全屏障,锆合金包壳材料的显微组织一直是国内外的研究热点。针对目前国内外在中子辐照对锆合金显微组织的影响研究领域现状进行了综述,总结了现有研究的不足并提出了对未来研究方向的建议：采用微观分析手段,针对不同中子注量下锆合金包壳的第二相、氢化物和氧化膜展开研究,获得其在实际使用工况下的演变规律,为我国锆合金包壳材料的优化改进提供支撑。     

基于GIS的低温冷冻灾害风险性评估

为了提高低温冷冻灾害的防御能力,为政府部门制定防灾减灾规划提供科学依据。本研究基于自然灾害形成机理及风险评估原理,以济南市长清区为例,利用气候资料、历史灾情数据、地理信息数据及社会经济等资料,采用层次分析法和加权综合评价法,借助GIS空间分析技术,通过对致灾因子危险性、敏感性、易损性和防灾减灾能力4个评价指标的空间评价,实现了对长清区低温冷冻灾害的风险性评估和综合区划。结果表明：长清区低温冷冻灾害总体表现为空间化强,没有规则的分布规律,综合风险性总体水平较高。文昌街道办事处、五峰山街道办事处、双泉乡的部分地区综合风险性最高,属于低温冷冻灾害的重点防御区域;轻、低风险主要分布在平安街道办事处、万德镇、马山镇、崮云湖街道办事处的部分辖区。该评估结果将对本地区农作物的种植规划及灾害防御措施的制定提供参考依据。     

新一代温室气体排放情景下安徽省未来气候变化预估分析

基于联合国政府间气候变化专门委员会(IPCC)发布的新一代排放情景,和中国气象局发布的"中国地区气候变化预估数据集V3.0",针对安徽省,笔者选择1971—2000年作为基准期,采用区域气候模式模拟的方法,对比分析安徽省未来气候变化特征,重点评估近期内即未来20年气候变化趋势。模拟结果显示,未来20年不同情景下安徽省平均气温均为上升,全省平均增温幅度约0.9~1.1℃;而降水量变化具有不同特征,较低排放情景下降水量以上升为主,高排放情景则以下降为主。到2050s全省平均气温相比于基准期将升高1.6~1.7℃,升温幅度呈现北高南低的特征;全省降水量相比于基准期将下降50~90 mm,各地降水量均呈下降趋势。另外,通过对降水和气温的模拟,预估未来该地区旱涝演替更加频繁,高温热浪等事件也将进一步频发。     

转基因抗虫玉米的定量检测

建立高效、快速、准确的抗虫转基因玉米MIR162转化体特异性实时荧光PCR(real-time PCR)检测方法。采用SYBR Green I和Taqman探针2种定量方法对抗虫转基因玉米MIR162进行了检测研究。SYBR Green I法MIR162转化体特异性片段和内标基因zSSIIb的熔解曲线均表现为单一峰,扩增特异性强;二者的扩增效率均在90%~100%之间,标准曲线相关系数R2均大于0.998。2种定量方法的检测限(LOD)均为24个拷贝,定量限(LOQ)为48个拷贝,灵敏度较高。重复测试结果表明,SYBR Green I法的标准偏差(SD)变化范围为0.05~0.13,相对标准偏差(RSD)范围为0.17%~0.40%,Taqman探针法的SD变化范围为0.03~0.14,RSD范围为0.11%~0.44%,说明2种定量检测方法具有良好的重复性和精密度。对MIR162含量为3%、1%和0.5%的混合样品的定量结果为：3.13%、1.09%、0.53%（SYBR Green I法）和3.07%、1.02%、0.50%（Taqman探针法）,RSD均小于10%,T检验分析表明2种定量方法定值结果差异不显著。本研究建立的2种实时荧光定量PCR均可有效对转基因玉米MIR162进行定量检测,可根据不同实际需求及实验室条件进行选择相应的检测方法。     

A dwarfing mutant caused by deactivation function of alpha subunit of the heterotrimeric G-protein in rice

Dwarf mutants in plants are crucial for elucidating regulatory mechanisms for plant growth and development. Previous studies suggested that the heterotrimeric G-protein alpha subunit known as D1/RGA1 in rice was involved in deactivation function of the G protein. However, so far no partner has been analyzed the spatial structure change acting with D1. In this study, a dwarf mutant designated Mu101 was obtained in M₂ population of rice indica cultivar M804 treated with ⁶⁰Co γ-ray. Genetic analysis of Mu101 indicated that the dwarf phenotype was controlled by a single dwarf gene, and the dwarf mutant was insensitive to gibberellin (GA), which was named dwarf 89 (d89). Using a large F₂ population derived from a cross between the d89 and a japonica rice variety, Taigeng16, the D89 gene was fine mapped into a 62.13 kb physical distance on chromosome 5, where eight open reading frames were predicted. Sequence analysis indicated that only one bp substitution (A-G) was found in LOC_Os05g26890 between M804 and the d89 mutant. The rice GA insensitive dwarf mutant DWARF1 gene was in this locus. The modeling analysis showed amino acid threonine to alanine mutation was likely to make the alpha helix short, and led to the G protein deactivation.     

油松端粒酶活性的测定方法

端粒酶具有修复延伸端粒的功能,对于维持真核生物染色体的完整性至关重要,因此对其活性的测定有着重要意义。通过大量的摸索实验,将常用于动物细胞端粒酶活性检测的端粒末端重复序列扩增法（TRAP法）运用到对植物材料的检测中来,对北方常见木本植物油松进行了端粒酶活性测定。基于TRAP法的原理,对体外延伸、引物选择、退火温度、电泳检测等实验条件进行优化。结果表明,油松端粒酶活性测定的最佳体外延伸条件为30℃延伸1 h,最适前导链引物和反向引物分别为TS21和ACX,PCR退火温度为61.8℃,电泳检测使用Gene Finder核酸染料染色效果更好,建立了一种有效测定木本植物油松松针叶端粒酶活性的测定方法。     

7种稀土小檗碱配合物对4种农作物病菌的抑菌性及急性毒理性的研究

研究开发了7种新型稀土配合物。使用差热热重和红外表征了配合物,采用最低抑菌浓度实验,研究了配合物对4种（番茄溃疡病菌、向日葵菌核病菌、尖孢镰刀菌和胡萝卜黑斑病菌）农作物病原菌的敏感性;采用急性经口毒性和急性经皮毒性试验,考察其急性毒性。7种配合物均为低毒性药物,并且抑菌效果好。对向日葵菌核病菌最低抑菌浓度均为0.125 mg/mL。对番茄溃疡病菌抑菌效果最好的是Ce(NO3)3(B)3、Gd(NO3)3(B)3,最低抑菌浓度是0.125 mg/mL。对尖孢镰刀菌抑菌性最好的是La(NO3)3(B)3、Gd(NO3)3(B)3和Y(NO3)3(B)3,最低抑菌浓度是0.5 mg/mL。对胡萝卜黑斑病菌抑菌性最好的是Gd(NO3)3(B)3,最低抑菌浓度是0.5 mg/mL。结果表明7种新型稀土小檗碱配合物是一类广谱、高效、低毒的杀菌剂。     

串珠镰刀菌的原生质体制备研究

通过观察离心所产生沉淀的松紧度、原生质体复苏的再生率、离心后液体的OD值来研究甘蔗梢腐病病原菌--串珠镰刀菌的原生质体制备技术。文中对液体高度、离心力、离心管大小、离心机升速与减速的快慢、不同洗涤剂等因素对原生质体活性、沉淀松紧度及壁效应的影响进行研究。对不同酶浓度及不同酶系组成对原生质体产率也进行了影响。此外,还介绍了一种离心管平衡的方法。研究结果认为：10 mL 0.1g/mL溶壁酶对0.5 g串珠镰刀菌菌丝进行裂解可以得到大量原生质体;进行裂解时使用溶壁酶与崩溃酶混合则可以获得更多的原生质体。以1 mol/L山梨醇或1.2 mol/L KCl洗涤沉淀并再离心, 使用底部较小的离心管和较少量的液体,并缓慢减速停止离心,有利于原生质体沉淀且能避免沉淀松散。