The expression of Quox-1 gene in the embryo development ofAndrias davianus

Two set ofAndrias davidanus fertilized eggs and its seedlings were got by means of artificial reproduction. The sections of the embryo ofAndrias davidianus were made by histological method. The expression of homologous sequences of Quox-1 gene has been studied by immunohistochemical method with special antibody. Results show that the expression of homologous sequences of Quox-1 gene was detected in the epidermis of blastula and gastula. The highly expression was investigated in the epidermis at the neural tube phrase and the tail bud stage. Deng Fengjiao: born in October, 1954, Associate Professor     

The development of embryo and endosperm inRanalisma rostratum

The embryo and endosperm development inRanalisma rostratum was studied in this paper. The zygote divides by a transverse wall to form basal and apical cells. The larger basal cell undergoes no further division and becomes the basal suspensor cell. The embryo development conforms to the Caryophyllad type. In the mature seed, the embryo is U-shaped and forms the embryonic shoot apex accompanied by one intraseminal leaf. The endosperm development belongs to the Hebolial type. The primary endosperm nucleus (invariably lying in the chalazal part of the embryo sac) divides and forms a large micropylar chamber and a small chalazal chamber. The chalazal endosperm chamber remains uninucleate, and free nuclear divisions occur in the micropylar chamber. Cellularization of the endosperm begins simultaneously from the micropylar and the chalazal ends. The cellularization is related to phragmoplast and cell plate, and does not arise from the free growth wall. Accompanying the embryo development, the endosperm degrades and does not exist in the mature seed. In the proembryo developing stages, the basal suspensor cell and chalazal endosperm cell grow to a considerable size. The nucleus and cytoplasm stain densely. Sometimes, binucleate or trinucleate was observed in both of the two cells. In the laterally concave embryo stage, they begin to degenerate which coincides with the cellularization of the endosperm. They are presumed to participate in the nourishment of the embryo proper in the early stages of embryo development. Supported by the National Natural Science Foundation of China Wang Jianbo: born in May. 1964, Ph. D.     

Expression ofChlamydomonas actin-gfp fusion gene in tobacco suspension cell and polymerization of the actin-gfp proteinin vitro

The fusion gene of actin (cDNA ofChlamydomonas reinhardtii) and green fluorescence protein (gfp) had been constructed into two expression vectors which could be expressed inE. coli and tobacco suspension cells BY2. The correct expression was observed inE. coli and BY2 with a fluorescence microscopy. The fusion protein, which took part in the membrane skeleton, was mainly located peripherally along the membrane, specially the fusion protein was distributed around nucleus and cell plate, while the fusion protein also forms F-actin in the cell. The fusion protein was purified from Bl21plus by ammonium sulfate fractionation, ion exchange chromatography and hydrophobic interaction chromatography. The purified production could polymerize into F-actin when the actin polymerizing buffer was added. It was demonstrated that the characteristics and function of actin inChlamydomonas was similar with those of animals and higher plants.     

稻瘟病抗性基因 Pi63 启动子4 个构建体的表达及抗性分析

利用农杆菌介导法获得带有不同长度启动子区域的稻瘟病抗性基因Pi63全长基因的转基因水稻,并采用实时荧光定量PCR方法鉴定了T1代阳性植株的基因拷贝数,进一步测定了单拷贝转基因植株在接种稻瘟病菌前后Pi63基因的表达量变化.结果表明:接种后4个分别带有不同长度启动子的Pi63转基因植物T1代中Pi63基因的表达量均升高,其抗病相关的顺式作用元件处于P1与P2中间的区域,P0到P1的区域存在负调控元件.此结果可为进一步揭示水稻稻瘟病抗性基因Pi63的分子机理奠定基础.     

The internal structure of Early Cambrian fossil embryo <Emphasis Type="Italic">Olivooides</Emphasis> revealed in the light of Synchrotron X-ray Tomographic Microscopy

Countless fossil embryo Olivooides and the hatched larvae, juveniles and adults （the latter two kinds are Punctatus） are recovered by means of acid maceration from the fine-crystalline to medium-crystalline phosphatic limestone and phosphatic micrite of Early Cambrian Kuanchuanpu Formation at the Shizhonggou section, near Kuanchuanpu Village, Ningqiang County, Shaanxi Province, China. Using the technique of Synchrotron X-ray Tomographic Microscopy, the 3D internal structure of Ofivooides and Punctatus is reconstructed. The morphological and statistic analyses are also given to the stellae structure of Ofivooides and Punctatus, which indicates that this structure is a result of adaptive evolution to a lifestyle of fast-attaching after hatching, probably with the function of mucilage secretion. The internal structure of Punctatus is described and discussed. The ovum-like structure, a common internal feature of Punctatus, is considered as the taphonomic structure, rather than eggs or other biological structure. This structure is thought to be formed after the burial of the animal and before or during the mineralization. The original internal structure of Punctatus is assumed to be tabulae-filled, with soft body grown on them.     

斜带石斑鱼ERP44基因的克隆与表达分析

为进一步了解内质网蛋白44基因(ERP44)在石斑鱼免疫反应中的作用,本研究根据实验室石斑鱼转录组数据中ERP44的表达序列标签(EST)设计引物,克隆了斜带石斑鱼(Epinephelus coioides)Ec-ERP44基因的开放阅读框序列(ORF),利用生物信息学手段分析Ec-ERP44基因及其编码蛋白的序列结构和特征,并通过实时荧光定量PCR检测该基因的组织分布特征,以及脂多糖(Lipopolysaccharide, LPS)、聚肌胞苷酸(Polyinosinic-polycytidylic acid, Poly I:C)刺激后该基因在石斑鱼脾脏中的表达变化。研究结果表明,Ec-ERP44基因ORF全长1 233 bp,编码410个氨基酸;该蛋白具有蛋白质二硫键异构酶(PDI)家族保守的硫氧还蛋白结构域。同时,Ec-ERP44基因在健康石斑鱼体内的多种组织均有表达,其中在脑组织中的表达量最高;在LPS与Poly I:C刺激后,石斑鱼脾脏中Ec-ERP44基因的表达显著上调。本研究结果表明,Ec-ERP44基因参与了石斑鱼抗病原感染的免疫反应。     

Cloning oflea cDNA fragment of carrot (Daucus carota L.) and analysis of its expression features

Addition of concentrated sucrose to MS culture arrests the development of carrot somatic embryo at the stage of cotyledon embryo and, with the sucrose concentration restored to normal level, the embryo thus arrested is reactivated into post-embryonic development. Using the method of RT-PCR, the cDNA fragment of a new member of the Dc3 family of lea has been obtained from carrot somatic embryo under regulated state. As revealed by Northern blotting, strong expression has been observed in carrot somatic embryo under regulated state but the expression was much reduced 12 h after deregulation, and nearly disappeared 24 h after. Based on this finding as well as results of related studies, it is surmised that changing the sucrose concentration in culture enabled carrot somatic embryo under suspension culture to undergo a specific course of development which is comparable to the dormancy-germination process of seeds.     

核桃JrEFM1转录因子响应逆境及激素的表达模式

MYB转录因子在调控植物生长发育和逆境响应方面发挥着重要的作用.通过克隆获得了核桃的1条R1-MYB类转录因子EFM基因(命名为JrEFM1),利用生物信息学和实时荧光定量RT-PCR(RT-qPCR)技术,分析在不同非生物及植物激素处理下JrEFM1的表达规律,探究了JrEFM1的基本生物学功能.结果显示,JrEFM1的编码区长为1 320 bp,编码蛋白含439个氨基酸,分子量为48.322 KDa,理论等电点为9.26.与葡萄、番薯等具有较近的进化关系.其启动子包含干旱胁迫(MBS)、热激响应(HSE)、水杨酸(SA)、玉米素(O2-site)和赤霉素响应(GARE-motif)等相关元件.对JrEFM1在干旱,冷害,热激,ABA,JA,SA处理下的表达情况进行分析,发现JrEFM1可被这些处理不同程度地诱导,同时根和叶表现出不同的转录水平.表明JrEFM1可响应逆境胁迫,并与激素信号通路相关;JrEFM1可作为核桃逆境响应机制研究及抗逆育种的优良候选基因.     

Analysis of IGF2 mRNA expression and its methylation status between cattle yaks and their parents

Cattle yaks are an F1 hybrid between cattle and yaks and exhibit significant hybrid vigor. However, cattle yaks are sterile males. To study the epigenetic aspects of this reproductive isolation, IGF2 mRNA expression in cattle yaks and their parents and the patterns of the IGF2 differentially methylated regions (DMR) were examined in blood and testes. The results showed that IGF2 expression in cattle yaks’ testes was lower than in their parents (P < 0.01). The IGF2 DMR was highly methylated (above 90%) in blood and testes of cattle yaks, yaks and cattle, with the highest level in cattle yaks (P > 0.05). Our study showed that IGF2 plays an important role in bovine spermatogenesis and might be involved in cattle-yak male sterility. The methylation level of the IGF2 DMR was irrelevant to the lower expression of IGF2 in cattle yaks; other factors perhaps play roles in its expression.     

控制水稻红米性状相关基因分子标记的开发

红米因其独特的营养价值日益受到人们关注.红米由两对非同源染色体上的Rc、Rd显性基因控制,且Rc、Rd基因同时存在时,水稻才表现为红米表型.本研究分别在Rc和rc等位基因以及Rd和rd等位基因影响性状的关键性位点上建立了分子标记:Rc(+5150)和Rd(+276).检测结果显示:这2个分子标记可以分别用来区别控制红米性状的Rc/rc和Rd/rd基因型.开展本研究,为今后利用分子标记辅助快速有效地选育各类红米水稻新品种研究提供了重要的帮助.     

苦荞CCT基因家族生物信息学及表达分析

CCT转录因子在调控植物花期、生长发育及抗非生物胁迫等方面发挥着重要的功能。本研究以拟南芥AtCCT基因家族为参考序列,利用本地BLAST并结合保守结构域等生物信息学工具,筛选出苦荞FtCCT基因家族成员,并对其理化性质、染色体分布、基因结构、系统进化及表达水平进行分析。结果显示:从苦荞中共鉴定出35个FtCCT基因,含1-8个内含子;编码蛋白有117-753个氨基酸残基,等电点为4.96-9.51,均为亲水性蛋白。染色体定位分析表明,这些基因在8条染色体上均有分布。苦荞FtCCT基因家族含有10个保守基序和5个保守结构域,且都含有CCT保守结构域。系统进化分析表明,苦荞的FtCCT基因家族与拟南芥一样可分为3个亚家族,其中CMF亚家族的成员最多。35个FtCCT基因在苦荞根、茎、叶和花中的表达水平具有差异性,在叶和花中具有高表达量的成员较多,只有少数的成员在根和茎中高表达。本研究为进一步解析CCT基因调控苦荞花期及生长发育奠定基础。     

Molecular cloning, identification and characteristics of NYD-SP9: Gene coding protein kinase presumably involved in spermatogenesis

Using cDNA microarray hybridization from a human testicular cDNA library, one gene exhibiting ten-fold difference at expression level between adult and embryo human testes was cloned and named NYD-SP9, which was believed to be involved in spermatogenesis. Southern blot hybridization results showed that NYD-SP9 expressed highly in testis but low in ovary. Protein motif analysis of this cDNA sequence revealed a cluster of phosphorylation sites, indicating its potential involvement in signal pathways during spermatogenesis. Furthermore, one transmembrane helix was predicted in N-terminal region, indicating that putative NYD-SP6 may be served as a transmembrane protein. The proximity of these potential phosphorylation sites to each other indicates that there may be interaction among these sites to regulate spermatogenesis. These findings suggested that protein kinase NYD-SP9 might play a role in male germ cell differentiation.     

菠菜HD-ZIP III基因家族鉴定与表达分析

植物叶片在生长发育过程中,近-远轴极性的建立与维持是叶片正常行使光合效应、呼吸作用、蒸腾作用的结构基础. HD-ZIP III是重要的叶片近轴极性基因,本研究在菠菜（Spinacia oleracea）基因组中鉴定到3个HD-ZIP III基因家族成员,依次命名为SoHB1,SoHB2和SoHB3,并对其蛋白理化性质、保守结构域和进化关系进行了分析. 研究结果表明：3个HD-ZIP III蛋白都具有4个保守结构域. 共线性分析表明：菠菜SoHB1,SoHB2和SoHB3分别与拟南芥ATHB8,ATREV和ATHB15存在共线性关系. 启动子顺式作用元件预测结果表明：菠菜HD-ZIP III家族基因启动子上游包含光响应、逆境胁迫反应、激素响应等顺式作用元件. 在不同叶型菠菜种质中的表达模式分析结果表明：单戟型叶片的SoHBs表达量最高;此外,经干旱胁迫处理后,SoHB1,SoHB2和SoHB3的基因表达量普遍下降;对菠菜包括根、茎、薹叶、功能叶、薹叶柄、功能叶柄等部位进行表达分析,发现SoHB1在根部表达最为丰富,SoHB2和SoHB3在薹叶处表达最为丰富.     

菠菜叶酸合成代谢途径基因鉴定及表达谱分析

从菠菜(Spinacia oleracea L.)基因组数据库中筛选鉴定了与菠菜叶酸合成转运,及与C1代谢相关的25个基因,并对其编码的蛋白做进化树和保守域分析,发现叶酸蛋白在进化上表现出保守型和复杂性。用实时荧光定量聚合酶链式反应(qRT-PCR)分析了叶酸含量不同的菠菜的叶酸相关基因表达量,发现在转录水平上只有少数叶酸基因表达量与菠菜叶酸含量有关。     

Characterization of the flagellar biosynthesis regulatory geneflbD inAzospirillum brasilense

A flagellar gene cluster fragment includingflbD ofAzospirillum brasilense was cloned and sequenced. TheflbD mutant strain was found to be nonmotile—losing both polar and lateral flagella (Fla⁻Laf⁻). Motility and flagella were regained by complementation with plasmid-borne multicopyflbD, but altered with larger swarming circle and fewer lateral flagella on the semisolid plate. This result indicated that FlbD plays an important role in the regulation of both polar and lateral flagellar biosynthesis inA. brasilense.     

Investisations into the perplexing interrelationship of the Genus Takifugu Abe, 1949 （Tetraodontiformes,Tetraodontidae）

The phylogenetic relationships within the genus Takifugu Abe, 1949 （Tetraodontiformes, Tetraodontidae） remain unresolved. Because of the use of Takifugu as model organisms, the resolution of these relationships is crucial for the interpretation of evolutionary trends in biology. Pufferfishes of this genus are comprised of a comparatively small number of species and are mainly distributed along the coastal region of the western part of the Sea of Japan and the coastline of China. Mitochondrial gene sequences were employed to test the phylogenetic hypotheses within the genus. Seventeen species of the genus were examined. Molecular phylogenetic trees were constructed using the maximum parsi- mony, neighbor-joining, maximum likelihood and Bayesian methods. Our hypothesis of internal relationships within the genus differs from previous hypotheses. Our results indicate that （1） the genus Takifugu is a monophyletic assemblage; （2） the genus is divided into 6 subgroups based on the moecular data; and （3） there is low genetic diversity among the species within this genus. In addition, speciation within Takifugu appears to be driven by hybridization and isolation by distribution. Our results also suggested that the taxonomy in the genus should be clarified based on both molecular and morphological data.     

枝状枝孢霉MD2的Unigene A03231的克隆及原核表达分析

为研究真菌紫杉醇合成相关基因的功能,从产紫杉醇真菌枝状枝孢霉MD2中克隆了Unigene A03231的DNA和c DNA全长序列(918 bp).结果表明:该基因不含内含子,其编码产物含305个氨基酸,与短链脱氢酶/还原酶一致性为64%,预测蛋白分子量为33071.5 Da,不存在信号肽,含有2个跨膜结构.Unigene A03231以单拷贝形式存在于基因组,在0~30 h内其表达水平随茉莉酸甲酯诱导表现为先升高后降低.构建了该基因的原核表达菌株,初步优化了其在大肠杆菌BL21中的诱导表达条件为:诱导温度28℃,IPTG浓度0.6 mmol/L,诱导时间8 h.为深入分析该基因的催化功能奠定了基础.