荔枝FKBP16-2基因启动子的克隆与瞬时表达分析

前期研究中发现了一个果肉低表达而叶片中高表达的荔枝基因FKBP16-2。本文克隆了该基因1 578 bp的启动子片段并对其功能进行了初步分析,结果表明:荔枝FKBP16-2基因启动子序列中含有大量的TATA-box和CAAT-box保守元件,以及TCA-element,ARE,HSE,GCN4_motif,O2-site等各种转录调控相关的顺式作用元件。该启动子能驱动GUS基因在荔枝的花、叶、根、果皮以及种子中表达而在果肉中不表达,表达具有组织特异性。     

柱花草叶肉原生质体提取与瞬时表达体系构建

柱花草(Stylosanthes guianensis)是一种重要的热区豆科牧草,为利用原生质体瞬时表达体系开展柱花草基因功能研究,以20 d龄期‘热研5号’柱花草子叶为试验材料,通过正交试验考察不同因素对原生质体分离及转化效率的影响。结果表明：采用1.5%纤维素酶、1.25%离析酶、0.5 mol·L^-1甘露醇和4 mmol·L^-1MES酶解液组合,酶解时间为16 h时,原生质体的产量和活性均达到最佳,分别为(4.567×10⁶)个·g^-1和92.271%;原生质体浓度为(6×10⁵)个·mL^-1、质粒浓度为0.6μg·μL^-1,PEG-4000浓度为50%、转化时间为5 min时,转化效率最高,可达52.524%。构建了目标蛋白SgRKL1表达载体pA7-BIUTNT::SgRKL1-GFP,利用PEG介导法将其转入柱花草原生质体,激光扫描共聚焦显微镜检测结果显示SgRKL1蛋白定位于细胞膜上,说明所建立的柱花草叶肉原生质体瞬时表...     

活塞组机械疲劳试验的有限元仿真

对某铝合金活塞在机械疲劳试验机上做的疲劳试验进行有限元分析,为活塞的改进设计提供依据。采用装有活塞销的三维整体活塞有限元模型,应用接触单元模拟实际情况,对活塞在交变机械载荷作用下的应力和应变进行了计算,找出了危险点部位,计算结果与试验吻合。     

管网中泵破坏瞬变流的数值模拟

建立了管网中泵破坏瞬变流的数值模拟，该模型考虑了泵破坏无量纲非线性边界条件，运用特征线法对方程进行了求解。经验证，证明模型是可靠的，可用于工程分析计算。     

汽油机HCCI燃烧过渡过程控制策略

利用可变气门机构使部分残余废气留在缸内,是实现汽油机HCCI燃烧的切实可行的方法,然而多气门参数的调节又给燃烧控制增加了难度.本文通过试验,研究了全可变气门机构中4个控制变量对汽油机HCCI燃烧过程的影响,并对过渡过程中发动机负荷相对于各控制变量的动态响应时间进行了比较.研究发现,实现HCCI燃烧过渡过程的快速响应,应当采用适当的控制参数的耦合策略.试验结果表明,进、排气门相位的对称性原则和气门升程的同步性原则可以简化耦合策略,并改善IMEP的过渡响应.这一结果为4VVAS-HCCI汽油发动机过渡过程控制策略的研究提供了参考.     

草莓果实外源基因瞬时表达系统的建立

将东北红豆杉紫杉烷13α-羟基化酶(Taxane13α-hydroxylase,13OH)基因全长cDNA正向插入到pCAMBI-A1304,构建其植物表达载体pCT13OH;将水稻小RNA169d(microRNA169d,miRNA169d)基因前体(precursor)全长DNA正向插入到pCAMBIA1305.1,构建其植物表达载体pCO169d。用电击法把它们导入根癌农杆菌GV3101,获得有关工程菌株。用1mL无菌注射器分别将这2种工程农杆菌悬浮液注射到草莓(Fragaria×ananassa)授粉后1周、2周、3周的果实中,发现授粉后2周果实适宜注射,可用于瞬时表达。对授粉后2周的果实进行不同注射部位、根癌农杆菌不同活化时期和根癌农杆菌悬浮液不同注射量试验,以蒂部注射、根癌农杆菌活化12h和0.5mL悬浮液注射量的效果最好,与结构基因13OH和调节基因miRNA169d相融合的GUS基因都得到表达,瞬时表达成功。     

农杆菌介导的半夏GUS基因瞬时表达

采用根癌农杆菌感染半夏无菌叶片、叶柄和愈伤组织,对GUS基因的瞬间表达进行研究,并分析了不同转化受体、感染时间、菌液浓度、共培养、预培养时间、菌种对GUS基因的瞬间表达的影响。结果显示,以半夏的无菌苗叶柄和愈伤组织,在OD值为0.2的EHA101或EHA105农杆菌液中感染15 min,叶柄暗处共培养3 d,愈伤组织2 d,能够得到较高的GUS基因瞬间表达。     

Roles of Root Aerenchyma Development and Its Associated QTL in Dry Matter Production under Transient Moisture Stress in Rice

Abstract

Enhanced aerenchyma development in rice under transient drought-to-waterlogged (TD-W) stress promotes root system development by promoting lateral root production. This study analyzed the quantitative trait loci (QTLs) associated with the plasticity in aerenchyma development under TD-W stress. A mapping population of 60 F₂ genotypes of chromosome segment substituted lines (CSSL) derived from CSSL47 and Nipponbare crosses were grown in rootboxes and evaluated for shoot and root growth, and aerenchyma development (expressed as root porosity). The TD-W stress was imposed starting with water saturated soil condition at sowing and then to progressive drought from 0 to 21 days after sowing (DAS) prior to exposure to sudden waterlogging for another 17 days (21 to 38 DAS). We performed simple and composite interval mapping to identify QTLs for aerenchyma development. QTL associated with aerenchyma development was mapped on the short-arm of chromosome 12 and designated as qAER-12. The effect of qAER-12 on the plasticity in aerenchyma development under TD-W was significantly associated with the increase in lateral root elongation and branching. This resulted in greater root system development as expressed in total root length and consequently contributed to higher dry matter production. This qAER-12 is probably the first reported QTL associated with aerenchyma development in rice under TD-W and is a useful trait for the improvement of the adaptive capability under fluctuating soil moisture conditions.     

Modulation of K+ channels by Ca2+ and pH in regulatory volume decrease

It has been widely appreciated that animal cells rely on the mechanism of regulatory volume decrease(RVD) after swell in a hypotonic environment. Activation of K channels is crucial in the process of self-protection. There are a characterized increase in cytoplasmic Ca and a decrease in pH in the process of RVD in many cell types. Ca entry via transient receptor potential(TRP) channel is crucial for the cytoplasmic Ca increase, which in turn induces the decrease in pH.The increase in cytoplasmic Ca and decrease in pH activate or inhibit the activity of K channel, respectively. In this review, the regulatory network at cellular level between cytoplasmic Ca and pH, and the modulation of K channels by Ca and pH in the process of RVD are discussed.     

农杆菌介导的烟草瞬时表达影响因素研究

通过探究根癌农杆菌(Agrobacterium tumefaciens)介导的烟草瞬时表达体系的影响因素,确定烟草瞬时表达的最佳条件,为目的基因的功能研究提供技术支持。利用马铃薯晚疫病抗性基因RB和致病疫霉菌效应基因Avrblb 1作为报告基因,采用农杆菌介导的注射渗透法,摸索农杆菌菌悬液OD600值和菌系(遗传背景)、烟草品种及其生长时期等因素,确定烟草高效的基因瞬时表达条件。研究结果表明:介导烟草基因瞬时表达的根癌农杆菌菌悬液适宜OD600值为0.3~0.8;在菌悬液OD600值为0.1及以上时,AGL1、GV3101和C58C1介导的报告基因瞬时表达效率相似,均引发较强的叶片坏死反应;在农杆菌菌悬液OD600值小于0.1时,AGL1介导的瞬时表达效率最高。测试的烟草品种都具备较高的瞬时表达效率,苗龄6~8周的烟草适合分析。通过测试农杆菌菌悬液OD600值和菌系(遗传背景)、烟草品种及其生长时期等因素,确定本氏烟和普通烟均可实现基因高效瞬时表达的条件。