两种质粒中gus基因在植物细胞和根癌农杆菌中的表达特性研究

以含有基因转化操作过程中常用的两种质粒载体pBI121和pCAMBIA2301的根癌农杆菌EHA105为材料,分别转化甜瓜子叶,应用组织化学方法检测了甜瓜子叶和子叶培养后的愈伤组织及根癌农杆菌菌液的瞬时转化效果,研究了两种不同的质粒载体上所含的gus基因在根癌农杆菌中和植物细胞中的表达特性。结果表明,不同质粒载体上所含的gus基因的表达特性不同,质粒载体pBI121上所含的gus基因既能在植物细胞中能表达,也能在根癌农杆菌细胞中表达,而质粒载体pCAMBIA2301上所含的gus基因能在植物细胞中表达,但是不能在根癌农杆菌细胞中表达。     

苜蓿花叶病毒外壳蛋白基因在白三叶中的表达及转基因植株的抗病性分析

以白三叶(Trifolium repens L.)的两个品种Irrigation和Huia的子叶为转化体，用根癌农杆菌(Agrobacterium tumefaciens)介导法将外源的苜蓿花叶病毒外壳蛋白基因转入白三叶，经过筛选、分化和再生，得到了具有卡拉霉素抗性的再生植株。对这些植株进行PCR、Southern和Northern分析，结果表明，外源目的基因已经整合到白三叶基因组中并且得到了表达。对Northern分析呈阳性的植株进行了抗病性检测，证明表达苜蓿花叶病毒外壳蛋白基因的植株病症减轻，发病率、病情指数及病毒积累量都明显低于对照，有的甚至不表现症状，达到了免疫的程度。     

表达dsRNA的转基因烟草能阻止烟草花叶病毒的侵染*

将烟草花叶病毒(Tobacco mosaic virus,TMV)的部分运动蛋白基因(AMP)构建成反向重复结构，插入植物表达载体pBin438上，通过三亲交配法导人根癌农杆菌(Agrobactetqum tumefaciens)菌株EHl05后通过农杆菌介导法转化烟草(Nicotiana tabacam)品种云烟87，获得了50株转基因烟草。PCR和Southern blot分析表明目标基因已整合到烟草植株的基因组内。用TMV对转基因烟草进行挑战接种，症状观察和病毒浓度的TAS-ELISA检测表明，转基因烟草对TMV的抗性包括3种类型：免疫型占10％；抗病型占4％；感病型占86％。Northern blot分析表明，目标基因mRNA在不同抗病类型植株中的积累量存在明显差异，目标基因mRNA的积累量与抗病性成负相关。     

根癌农杆菌介导水稻基因转化研究进展

简要综述了根癌农杆菌介导水稻基因转化的优点及转化后代的遗传学行为 ,着重分析了影响农杆菌介导水稻基因转化的各种因素和存在问题     

马铃薯Y病毒CP基因RNAi载体构建与干涉效果测定

RNA沉默（RNAi）介导的植物抗病毒研究是近年来引起广泛关注的一项植物抗病毒基因工程策略。马铃薯Y病毒（PVY）在世界范围内引起马铃薯、烟草等重要经济作物的病害,并且造成严重的经济损失。本文以PVY的外壳蛋白（CP）基因为模板扩增300bp和354bp两个片段,正向和反向分别插入植物表达载体pROKⅡ的35S启动子下游,从而构建了以PVY的CP基因为靶标的RNAi植物表达载体pROKY300,转入农杆菌EHA105中,以农杆菌渗入法在本氏烟中瞬时表达与wyCP同源的hairpin RNA。结果表明,瞬时表达的hairpin RNA有效干涉了PVY侵染。     

甘露糖阳性选择系统的建立及在番茄转化中的应用

利用PCR克隆了大肠杆菌(Escherichia coli)6-磷酸甘露糖异构酶基因（pmi)并进行了序列测定。将该基因替换植物表达载体pAMBIA2301中的卡那霉素抗性基因，构建了以pmi为选择标记基因的植物表达载体pPMI，并导入根癌农杆菌(Agrobacterium tumefaciens)EHA105中。研究了甘露糖对番茄(Lycopersicum esculentum)生长和生根的影响及叶盘对甘露糖的敏感性。通过根癌农杆菌介导对番茄进行遗传转化，获得转化番茄再生植株，并对转化植株进行了PCR扩增检测。研究表明以甘露糖阳性选择系统在番茄遗传转化中应用是可行的。     

反义乙烯受体LeFTR2基因对番茄的转化和功能分析

采用根癌农杆菌(Agrobacterium tumefaciens)介导法将LeETR2的部分反义序列导入番茄(Lycopersicon esculentum)子叶外植体，经抗性筛选和组织培养获得再生植株。通过对转基因植株中插入序列的PCR检测和Southern杂交，证明转化番茄成功。通过对LeETR2的功能的初步研究结果表明，转基因植株顶端优势丧失、极端矮化和番茄果实的内源乙烯合成量增加。这提示此基因在植物乙烯信号感受与转导系统中起着负调控的作用。     

Pt-mVirD2-GUS质体蛋白编码基因的合成及转化烟草的研究

将拟南芥冷调节基因AtCor15A的质体定位信号肽编码序列、丧失了核定位信号序列的根癌农杆菌VirD2基因突变序列mVirD2和GUS基因序列,拼接成pt-mVirD2-GUS嵌合基因,由CaMV 35S启动子和Tons终止子控制,并经根癌农杆菌介导,成功导入了烟草基因组.     

花青素调节基因Lc对转基因烟草和矮牵牛花色的影响

利用PCR的方法从油菜(Brassica napus)基因组中扩增得到花瓣特异性启动子XY355，将XY355与玉米花青素调节基因Lc融合构建植物表达载体pXY60。用冻融法将pXY60转入根癌农杆菌（Agrobacterium tumefaciens）LBA4404，然后以烟草（Nicotiana tabaccum L.）和矮牵牛（Petunia hybrida）的叶盘为外植体，通过根癌农杆菌介导法进行遗传转化。对再生植株的PCR检测证明外源基因已经整合到转基因烟草和矮牵牛的基因组中。对转基因烟草和矮牵牛的表型进行观察：转Lc基因烟草的花色由浅红变成了红色，转Lc基因矮牵牛的花色由白色变成了浅紫色。     

芪合酶基因转化小白菜的研究*

利用质粒pBin438构建了含有NPT Ⅱ基因和葡萄芪合酶(RS)基因的芪合酶组成型植物表达载体EHA105/pBinRS。用根癌农杆菌(Agrobacterium tumefaciens)介导法将RS基因导入纯合系小白菜(Brassica campestris ssp. chinensis)品种中，经Kan抗性筛选、PCR及RT-PCR检测，RS基因已整合到小白菜基因组中并得到了表达，共获得了7株转基因植株。     

Heterogeneity of Organic Matter of Aggregates of Protocalcic Chernozems

Eurasian Soil Science - Layers were step-by-step removed from macroaggregates (2–1 mm in diameter) of Protocalcic Chernozems via successive abrasion in a revolving rotator during 5, 10, 15,...     

食用仙人掌萎蔫气象条件分析

塑料大棚内种植的食用仙人掌在土壤墒情较好时也有萎蔫现象发生，通过试验观测和对仙人掌生理习性的分析，发现阴雨过后天气突然放晴温度急剧上升易使仙人掌发生萎蔫现象，并提出了田间管理的应对措施。     

姬松茸多糖提取工艺的优化

对姬松茸子实体多糖提取的工艺条件中的多糖提取温度、提取时间、浸提液pH值3因子的最优化组合问题进行了定量研究,建立了具有良好预测性能的姬松茸多糖提取条件的模型,并利用回归模型对工艺条件的最优化组合,对各单因子要素的多糖得率及其交互作用进行了探讨。试验表明,当浸提温度为100℃、浸提时间为3h、浸提液pH值为6.3时多糖得率处于较高水平     

济南山丘区土壤侵蚀潜在危险度景观格局分析

土壤侵蚀潜在危险度景观格局的形成及其变化是自然的和人为的多种因素相互作用的结果 ,运用景观生态学原理 ,通过分析景观斑块的类型、数量、面积大小和空间组合状况 ,揭示出济南市山丘区的土壤侵蚀潜在危险度分布特征及空间变异规律     

青海省耕地资源开发利用分析及对策研究

分析论述了青海省耕地资源开发利用的现状、特点和问题。在此基础上,提出了对青海省耕地资源进行研究的框架体系和思路,同时基于GIS/RS技术设计了相关的技术路线。最后依据所做设计对青海省耕地资源开发利用做了初步分析,并进行了相关的对策研究分析。     

Physical Properties of the Soils of Sibay City of the Republic of Bashkortostan

Eurasian Soil Science - The results of the study of the properties of urban soils of the city of Sibay located in the mining region of the Republic of Bashkortostan are presented. A specific...     

Characterization of physical and chemical properties of Varzea soils of Goias State of Brazil

Abstract

Brazil has approximately 30 million hectares of lowland areas, known locally as “Varzea,”; distributed throughout the country. Soils in these areas have the potential to support agricultural production, but very little is known about their fertility. The current experiment was undertaken to characterize the chemical and physical properties of representative “Varzea”; soils collected at 0–20, 20–40, 40–60, and 60–80 cm depth intervals from 23 sites in Goias State of Brazil. Organic matter contents averaged 42 g/kg in the surface 20 cm of soil and should make a significant contribution to overall nutrient availability. Soil pH increased slightly with depth from a mean value of 5.2 in the surface 20 cm of soil to a mean value of 5.4 in the 60–80 cm depth interval. Several soils had exchangeable Al values ≥ 1.0 cmol Al/kg, but soil Al saturation was generally less than the 60% level frequently associated with Al toxicity. However, cereal and legume production could benefit from lime addition in many of these soils. Although base saturation was fairly low in some soils, exchangeable Ca and Mg levels were, in general, adequate throughout the profile. Extractable P levels were adequate in most surface soil samples. Extractable K levels in these soils were generally low, and the application of K fertilizers should be beneficial, especially in conjunction with lime addition. The high clay content “Varzea”; soils of the Brazilian lowlands have some drainage problems but generally exhibit favorable chemical properties for crop production when compared to soils of the well‐drained Cerrados.     

Estimation of potential nitrogen-fixing activity of agrophytocenoses soils of the subtropical zone of Russia

An estimation of potential nitrogen-fixing activity of various soil types under agrophytocenoses of tea, filbert, peach, and pawpaw which grow in the area of Greater Sochi is given. High indicators of nitrogen fixation activity in the soil in the course of vegetation were characteristic of agrocenoses of pawpaw, peach, and filbert. The lowest level of nitrogen fixation was noted in the soil of a tea plantation. A negative correlation dependence of potential nitrogen-fixing activity on the acidity of the soil solution was established. A study of the seasonal dynamics of the activity of nitrogen fixation of soils of agrocenoses of the subtropical zone of Russia made it possible to distinguish a less active winter and more active spring-summer-autumn periods.     

淤地坝坝体体积的计算

对于已确定的淤地坝坝址,坝体的纵断面（即沟道横断面）是一定的。根据实测的沟造横断面,以沟底最低点为坐标原点,沟道一侧的岸壁曲线可用幂函数形式表示之。利用这种幂函数关系,对于坝坡均一的坝体．在设定坝顶宽、坝高和上下游边坡之后,作者推导出沟道一侧坝体体积的计算公式;沟道两侧的坝体体积即为坝体总体积。对于非均一坝坡、设有马道的坝和坝体加高的体积,可将坝体横断面分割为由几个均一坝坡组成的坝体断面,分别计算其体积。这种计算方法的精度,取决于坝址横断面岸壁曲线回归方程与实际岸壁的符合程度。