sch蚕胚胎期温敏性的mRNA差异显示研究

采用mRNA差异显示技术对不同催青处理的伴性赤蚁sch系统进行研究 ,并以普通黑蚁系统夏芳作为对照。实验中分别抽提高温和常温催青处理 36h后的sch蚕卵和夏芳蚕卵的总RNA进行DDRT -PCR分析 ,其产物于变性聚丙烯酰胺凝胶上电泳展示 ,得到大量的差示片段。对高温催青处理后在夏sch蚕卵中特异表达的 2条能稳定重复的带进行了克隆 ,得到了长度分别为 2 73和 199bp的cDNA片段 ,并对其进行了序列分析。Northern杂交证实HAP82 80仅在高温催青处理的夏sch蚕卵中有转录 ,表明其为sch蚕卵经高温催青处理后特异表达的产物     

实木橱柜的包装设计

实木橱柜体积大、质量重,通常采用整体包装的方式。根据物流过程情况提出半封闭和全封闭2种包装方式,并对全封闭包装中的瓦楞纸箱包装和蜂窝纸箱包装进行分析和比较。     

大豆孢囊线虫4号生理小种侵染大豆根系诱导表达的cDNA分析

大豆孢囊线虫(Heterodera glycines Ichinohe；Soybean Cyst Nematode，SCN)是一种土传的专性内寄生线虫。SCN的二龄幼虫侵入到大豆幼嫩的根组织中，导致大豆根内的细胞变形并与之形成“合胞体”。合胞体在形态上和生理上的变化是SCN直接诱导大豆基因表达的结果。本研究以高抗SCN的灰布支黑豆为材料，用大豆孢囊线虫二龄幼虫直接接种大豆的根系，应用DDRT—PCR技术及RDB(Reversedot—blotting)杂交鉴定，获得6个阳性cDNA克隆，分别是SCN侵染后5天的A32克隆(GenBank登录号为B1173978)；侵染后10天的B12克隆(GenBank登录号为B1173979)、B71克隆(GenBank登录号为B1173980)；侵染后15天的Cll克隆(GenBank登录号为B1173981)、CPl2(GenBank登录号为B1173982)克隆和CP32克隆(GenBank登录号为B1173983)。序列的同源比较表明，6个cDNA均与Shoemaker构建的大豆基因表达库中的cDNA序列有非常高的同源性，证明这些cDNA是大豆基因表达的产物。其中A32克隆的序列与控制拟南芥下胚轴生长的MYB转录因子、营养元素缺失诱导的番茄根的表达文库中的一个cDNA及番茄抗假单胞杆菌表达文库中的一个cDNA有较高的同源性。     

Simple sequence repeats as useful resources to study transcribed genes of cotton

Microsatellites or Simple Sequence Repeats(SSRs) are informative molecular genetic markers in many crop species. SSRs are PCR-based, highly polymorphic, abundant, widely distributed throughout the genome and inherited in a co-dominant manner in most cases. Here we describe the presence of SSRs in cDNAs of cotton. Thirty one SSR primer pairs of 220 (∼14%) tested led to PCR amplification of discrete fragments using cotton leaf cDNA as template. Sequence analysis showed 25% of 24randomly selected cDNA clones amplified with different SSR primer pairs contained repeat motifs. We further showed that sequences from the SSR-containing cDNAs were conserved across G. barbadense and G. hirsutum, revealing the importance of the SSR markers for comparative mapping of transcribed genes. Data mining for plant SSR-ESTs from the publicly available databases identified SSRs motifs in many plant species,including cotton, in a range of 1.1 to4.8% of the submitted ESTs for a given species. This revised version was published online in August 2006 with corrections to the Cover Date.     

New strategies for increasing heterozygosity in crops: Vicia faba mating system as a study case

Our research assesses the feasibility of using artificial selection on pre-mating floral traits to modify the mating system of faba bean (Vicia faba). This analysis considered two synthetic populations, which were derived from different genetic pools and had undergone five years of multiplication. For these populations, we identified floral and inflorescence traits that influence outcrossing per plant and examined the relative importance of these traits in governing yield. Codominant isozyme loci and the mixed-mating model were used to estimate the multilocus female outcrossing rate. A maternal half-sib design was used to evaluate the additive genetic component of floral and inflorescence traits, yield and yield components. Multiple regression was used to assess the effects of floral and inflorescence traits on outcrossing and yield and components of yield. The two populations exhibited mixing mating. Self-fertilization appears to result from the action of pollinating bees, so that its incidence could be modified by selection on floral and inflorescence traits that affect pollination. Floral and inflorescence traits affected individual differences in outcrossing unequally, with most variation being associated with the numbers of displayed flowers and inflorescences. Variation among plants in reward traits and in shape, although statistically significant, had limited and inconsistent influences on individual differences in outcrossing. Yield and its components varied strongly with aspects of floral display and, to a lesser extent, floral design, except for seed weight. Overall, our results imply that both outcrossing and yield could be enhanced by selection for plants that produce more inflorescences, each with relatively few flowers.     

水稻极度分蘖突变体的分离和遗传初步研究

通过EMS诱变处理栽培稻粳灿89(Oryza sative L.indica JX89）种子，从M2代植株中筛选得到一株突变体，其植株形态表现为叶细、色淡、植株矮化和极度分蘖。在将近一年的营养生长过程中，分蘖总数达到2000多个，命名为极度分蘖突变体（Excessive tillering mutant）ext37.ext 37自交所得M3和M4代植株表现同一表型。突变体自交及其与JX89和丰矮占5号（FAZ-5）发杂交F1、F2代结果表明该突变为显性遗传方式，可能涉及两对等位基因。为了观察突变基因对下游基因表达影响的程度，取生长期44天的亲本JX89和突变体ext37的节间分生组织提纯细胞总RNA，进行基因表达谱差异显示分析。结果表明突变体与亲本的基因表达模式有明显的差异，说明可能是处在上游调控基因的突变影响了下游许多基因的表达。目前有关分子生物学的进一步研究正在进行中。     

mRNA差异显示法分离与水稻籼粳亚种间杂交稻低温敏感不育性相关的cDNA片段

采用mRNA差异显示法对籼粳亚种间杂交稻亚优2号幼穗发育基因在两种不同温度条件下的表达差异进行了研究。结果发现,对照(正常温度28～30℃)和低温处理(白天24℃、夜晚20℃)的幼穗发育基因在转录水平上存在差异。从获得的35条差异片段中随机选取6条进行克隆测序,结果发现其中有1条片段长为556bp,暂命名为DF9,该片段与水稻第一染色体上一段cDNA序列有99%的同源性;其他5条片段与水稻的EST存在一定的同源性。进一步研究发现,DF9可能编码的产物与一些参与信号传导或能量代谢过程的酶存在不同程度的相似性。推测其与水稻籼粳亚种间杂种低温敏感不育性有关。     

LPG库的设计与运营

城市选用ＬＰＧ作燃料，有利于环境保护，方便居民。当前我国沿海地区城市采用ＬＰＧ作为燃料的热潮正迅猛发展。ＬＰＧ由丙烷和丁烷组成，二者一般采用分别储存方式，储存方式又分为常温带压储存和低温常压储存，分别介绍了这两种储存方式的设计条件。     

运用差异显示技术分离水花生在干旱胁迫下表达上调的基因

干旱发生时,水花生的须根将转变成又长又粗的肉质根,这有利于保水和从土壤深处汲取水分以减轻旱害.本研究中采用差异显示技术克隆干旱发生时与水花生根形态发生相关的基因,为进一步通过转基因提高农作物抗旱性奠定基础;共使用15对引物组合,回收到20个可能的在干旱诱导后表达上调的带,其中有一个能被反向northern所证实;而后对其进行了克隆和测序;同源性比对表明它可能是一个新基因的片段.半定量RT-PCR分析进一步证明该基因在干旱和盐胁迫下表达上调.     

mRNA差异显示技术在分离水稻抗性基因中的应用

mRNA差异显示技术从问世至今,在分离水稻抗性基因中得到了广泛的应用,通过对差异表达基因进行分析,能够进一步阐明水稻的抗性机理.对近年来mRNA差异显示法分离水稻抗性基因的研究做了总结,并从引物改进、PCR退火温度、dNTP浓度的改进、胶浓度的选择、标记方法的改进和差异片段验证方法的改进7个方面对mRNA差异显示技术的优化做了简单阐述.