小麦中一类依赖于NAD的细胞质型苹果酸脱氢酶cDNA的克隆和进化树分析

苹果酸脱氢酶普遍存在于各种生物中，它负责催化草酰乙酸和苹果酸之间的相互转换。根据其辅酶的特异性和在细胞内的分布和生理功能的不同，苹果酸脱氢酶在高等植物中可以区分出不同的类型。依赖于NAD的细胞质型苹果酸脱氢酶(cyMDH)是其中研究较少的一类。报道了从小麦(Tritium aestivum L.)中克隆的一种cyMDH的部分cDNA序列，并命名为W-MDH1。进化树分析表明来自不同生物的MDH可能是通过基因复制形成的。W-MDH1刖在小麦的根、茎、叶中呈组成型表达。     

酒类酒球菌苹果酸-乳酸酶基因mleA的克隆

苹果酸-乳酸酶是进行MLF的关键酶.该研究以酒类酒球菌31DH(Oenococcus oeni 31DH)的基因组DNA为模板进行其苹果酸-乳酸酶基因mleA的PCR扩增.PCR引物为5'-CGGAATTCATGACAGATC-CAGTAAGTAT-3'和5'-TAGGTACCACACTCTCAACACTCGTAAT-3',引物的5端分别引入EcoRI和KpnI酶切位点.得到的PCR产物约1.6kb.PCR产物回收后用EcoRI-KpnI双酶切,与经同样双酶切的质粒YEp352(大肠杆菌-酵母穿梭载体)进行连接并转化大肠杆菌感受态细胞,筛选重组质粒并用酶切及PCR验证.获得的酒类酒球菌苹果酸-乳酸酶基因的重组质粒命名为pLmleA.     

中华蜜蜂和意大利蜜蜂蜂毒磷脂酶A2基因的克隆与序列分析

从中华蜜蜂（Apis cerana cerana)和意大利蜜蜂（Apis mellifera)工蜂毒腺中快速抽提总RNA，用RT-PCR方法分别扩增各得到约含470bp的cDNA片段，将这2个片段克隆入PGEM-T载体，进行测序和序列分析，结果表明，这2个片段均含有405bp、编码蜂毒PLA2成熟肽的cDNA，并分别由cDNA推导出两者所编码的氨基酸序列，经序列比较，中蜂PLA2与意蜂PLA2具有95%的同源性，与大蜜蜂PLA2、美国熊蜂PLA2及黑腹果蝇PLA2的同源性分别为90%、54%、39%。     

节瓜CqWRKY1基因的遗传转化

为建立和优化节瓜的遗传转化体系,并高效地完成CqWRKY1基因的遗传转化,本研究以节瓜A39为材料,通过RT-PCR技术克隆了节瓜CqWRKY1基因,采用Gateway同源重组法构建了pEarleyGate101-CqWRKY1超表达载体,以节瓜子叶节为外植体,利用农杆菌介导的遗传转化法进行转化。结果表明,子叶节不定芽诱导的最适培养基为MS+0.5 mg·L~(-1)6-BA+1 mg·L~(-1)ABA,诱导率可达76.17%;生根的最佳培养基为1/2MS+0.5 mg·L~(-1)IAA;农杆菌侵染的最佳条件为农杆菌浓度OD_(600)为0.3、侵染时间为10 min、共培养时间为3 d;最佳的抑菌抗生素为500 mg·L~(-1)的头孢霉素。本研究在181株拟转基因组培苗中共得到76株阳性T_0转基因组培苗,其中共有37株移栽成活。PCR结果表明,CqWRKY1已成功整合到37株转化植株的基因组中。本研究结果为进一步研究CqWRKY1基因的功能奠定了一定的理论基础。     

胸腺肽基因对胡萝卜的遗传转化

胸腺肽(thymosin)是一种由胸腺分泌的促进淋巴细胞合成和成熟的多肽激素,对于淋巴系统的发育和维持免疫系统的平衡起重要作用(李金屏,1999;张符光和刘佃辛,1996)。市场上广泛使用的胸腺肽主要是从小牛胸腺和猪胸腺中提取制备的。从小牛胸腺和猪胸腺中进行胸腺肽的提取,生产成本     

青枯菌II型分泌系统编码基因的克隆

植物青枯菌II型分泌系统与致病蛋白的分泌密切相关。编码青枯菌II型分泌系统的gsp基因簇由12个基因组成,我们通过设计适合高GC%含量基因扩增的PCR反应体系,成功克隆了编码青枯菌II型分泌系统的全部12个gsp基因。分别将gsp基因连接到酵母双杂交系统的诱饵及捕获载体上,构建了gsp基因诱饵库和捕获库。经序列测定证明12个gsp基因读框正确,保障了其在酵母系统中的表达,为GSP蛋白之间的互作研究奠定了基础     

中西方风景园林的差异及其产生原因论述

风景园林艺术反映了人们的生活状态、对未来的憧憬以及文化程度,在一定程度上反映了一个国家人们的生活品质;同时,人们又将自身对于生活的向往和对未来的憧憬寄托在园林建造当中。所以,风景园林的建筑是物质和精神的融合,且每个国家建造风格也都各有不同。基于此,通过对中西方风景园林建筑进行分析,论述两者之间的差异以及产生差异的原因。     

吉富罗非鱼雌雄群体遗传差异的SSR分析

为探讨吉富罗非鱼（genetic improvement of farmed tilapia,GIFT）雌、雄群体间的遗传差异,本研究对国家级广西南宁罗非鱼良种场雌、雄吉富罗非鱼进行了遗传差异分析。研究结果表明,选取的11对SSR引物中有10对能获得稳定的目的条带;每个SSR基因座的等位基因数在2～4个之间,雌性罗非鱼的平均等位基因（Na）2.9个,稍高于雄性的2.8个;雌、雄吉富罗非鱼平均观察杂合度（HO）分别为0.4183和0.4154,多态信息含量（PIC）分别为0.4048和0.3932,属中度多态;雌雄个体间的遗传距离和相似性指数分别为0.0908和0.9132。此外,SSR基因座PRL-SO2在雄鱼中偏离Hardy Weinberg平衡（P〈0.005）。上述结果表明,吉富罗非鱼雌、雄群体的SSR多态性基本相同,推测这两者基因组间的差异较小。     

小白菜硝酸盐含量基因型差异的遗传行为研究

对硝酸盐含量有明显差异的2个小白菜品种江苏矮脚黄(P1)、上海青(P2)及其杂交一代(F1)、杂交二代(F2)和回交一代(B1、B2)6个群体共计460株进行了硝酸盐含量的检测,并采用联合尺度检验法分析其硝酸盐含量性状的遗传规律。结果表明,小白菜硝酸盐含量属于数量性状,其遗传表现符合加性 显性遗传模型,小白菜高硝酸盐含量对低硝酸盐含量性状为部分显性,显性度为0.45。通过对遗传力的估计值计算结果表明,小白菜硝酸盐含量广义遗传力为56.1%,狭义遗传力为21.6%,显性作用和环境影响都较大。因此,选育低硝酸盐含量的品种,应注重在高世代(如F5或F6)进行选择,不宜采用个体选择。     

两蜂种王浆蛋白质组分比较分析

王浆高产蜜蜂(Apis mellifera L.浆蜂)是我国20世纪90年代从意大利蜜蜂培育成功的新蜂种,产浆性能明显高于未经选育的意大利蜜蜂.中华蜜蜂(A. cerana cerana F.中蜂)是我国土生土长的蜂种,其蜂王浆产量低于以意大利蜜蜂为代表的西方蜜蜂.两蜂种所产蜂王浆的蛋白质组研究还未见报道.本研究利用双向电泳技术对浆蜂和中蜂新鲜蜂王浆全蛋白质组进行分析比较,为鉴定和判断蜂王浆的来源和蜂王浆功能成分的开发利用提供科学依据.     

Fatty Acid Inheritance in Wide Reciprocal Oilseed Crosses (Brassica rapa and B. napus)

Abstract

Erucic acid content and inheritance were studied in Brassica alboglabra, B. rapa and resynthesized B. napus from these parental species. Although significant variation in erucic acid content was found between the 20 B. alboglabra lines, all of them had to be classified as high erucic types. No major erucic acid gene polymorphism was identified.

In B. rapa the erucic acid content was regulated by one major gene, which showed partial dominance in three of the four crosses analysed. The high erucic gene seemed to function more efficiently in the Yellow Sarson cytoplasm, resulting in maternal differences when reciprocal crosses were compared. In crosses where modern low erucic acid lines were used as female parents the zero erucic acid F₂-individuals occurred more frequently than expected.

Segregation of the erucic acid content in crosses involving resynthesized B. napus showed a good fit to a two locus model. No maternal effects were found, but deviations from mid-parent values in the high erucic acid direction occurred in most of the F₁-hybrids, indicating partial dominance. Crosses between resynthesized oilseed rape and natural oilseed rape resulted in oleic acid transgressive F₅-lines.     

Proteomic analysis of royal jelly from three strains of western honeybees (Apis mellifera)

To compare the protein complement of royal jelly (RJ) from high RJ producing honeybees ( Apis mellifera L.), a strain of A. mellifera artificially selected for increased RJ production from Italian honeybees in China for more than two decades was compared to those of native Italian honeybees ( A. mellifera L.) and Carnica honeybees ( A. mellifera C.); the protein in RJ from these three strains of honeybees was partially identified by using a combination of two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF/MS), and a protein engine identification tool applied to the honeybee genome. The results showed that 152, 157, and 137 proteins were detected in the three species of RJ; among which 57, 57, 51 high abundant proteins ere identified, respectively. Most identifited spots, 45, 45, 41, were assigned to major royal jelly proteins (MRJPs). Remarkable differences were found in the heterogeneity of the MRJPs, in particular, MRJP3. Also, 3-glucose oxidase, 1-peroxiredoxin (PRDX), and 1-glutathione S-transferase (GST) S1 were identified in three RJ samples. Furthermore, during the determination of the peptides mass fingerprinting (PMF) of each spot, for the first time, PRDX and GST S1 proteins have been identified in RJ. Thus, the results suggest that the protein complement of high RJ producing honeybees is not different compared to native Italian honeybees, while a difference remains between Carnica honeybees.     

Characterization of royal jelly proteins in both Africanized and European honeybees (Apis mellifera) by two-dimensional gel electrophoresis

In this study, the proteins contained in royal jelly (RJ) produced by Africanized honeybees and European honeybees (Apis mellifera) haven been analyzed in detail and compared using two-dimensional gel electrophoresis, and the N-terminal amino acid sequence of each spot has been determined. Most spots were assigned to major royal jelly proteins (MRJPs). Remarkable differences were found in the heterogeneity of the MRJPs, in particular MRJP3, in terms of molecular weights and isoelectric points between the two species of RJ. Furthermore, during the determination of the N-terminal amino acid sequence of each spot, for the first time, MRJP4 protein has been identified, the existence of which had been only implied by cloning of its cDNA sequence. The presence of heterogeneous bands of glucose oxidase was also identified. Thus, the results suggest that two-dimensional gel electrophoresis provides a suitable method for the qualitative analysis of the proteins contained in RJ derived from different honeybee species.     

复杂疾病遗传基础及易感基因研究的发展趋势

复杂疾病如高血压、2型糖尿病以及恶性肿瘤等严重影响着人们的生活质量,并制约着社会经济的发展。因此,目前有关复杂疾病的项目攻关已成为学术研究的重点。以单核苷酸多态性以及拷贝数变异为基础的全基因组关联研究方法已日趋成熟。高通量测序技术的发展以及其成本的逐渐降低为更高精度的易感基因定位提供契机。本文总结了复杂疾病的遗传基础以及发病基础理论的研究现状,并对已经成熟的以及未来可预见的复杂疾病易感基因的定位及研究方法加以阐述和展望。     

Genetic Differences in Resistance to Iron Deficiency Chlorosis in Peanut

Iron (Fe) deficiency has been a widespread problem in peanut (Arachis hypogaea L.) grown on calcareous soils of northern China and has resulted in significant yield losses. Field observations showed considerable variability in visual chlorosis symptoms among peanut cultivars in the same soil. The objective of this study was to confirm the genetic differences in resistance to Fe-deficiency chlorosis in peanut and to identify feasible indicators for screening Fe-efficient genotypes. Resistance to Fe chlorosis of sixteen peanut cultivars grown on calcareous soil was evaluated in the field and physiological responses to Fe-deficiency stress were studied in nutrient solution. There were significant differences in resistance to Fe-deficiency chlorosis among the sixteen peanut cultivars tested, which was identified with SPAD readings, active Fe concentrations in young leaves in the early growth stages, and the pod yield. For Fe-resistant peanut cultivars, Fe-reduction capacity and quality of releasing hydrogen ions from roots increased under Fe-deficiency stress. Highly correlated relationships were observed between the summation of root Fe reduction and field chlorosis scores for sixteen cultivars (r² = 0.79). It was concluded that Fe-reduction capacity was a better physiological indicator for screening Fe-efficient peanut genotypes of the mechanisms measured.