An eight-nucleotide sequence in the potato virus X 3' untranslated region is required for both host protein binding and viral multiplication.

Gel retardation and UV-cross-linking techniques were used to demonstrate that two tobacco proteins, with approximate molecular masses of 28 and 32 kDa, bind to a site within the 3' region of potato virus X (PVX) genomic RNA. The protein binding is specific, in that a 50-fold excess of unlabeled probe prevents formation of the complexes but no reduction is observed with a 2,000-fold molar excess of yeast tRNA. Complex formation is inhibited by poly(U) but is relatively unaffected by poly(A), poly(G), or poly(C-I). PVX RNA-host protein complex formation occurs in vitro at salt concentrations up to 400 mM. Deletion mapping indicates that the proteins bind within the 3' untranslated region (UTR) of PVX genomic RNA and that an 8-nucleotide U-rich sequence (5'-UAUUUUCU) is required for the binding. Deletion of the 8-nucleotide U-rich region from the 3' UTR of a sensitive PVX reporter virus that carries the luciferase gene in place of the PVX coat protein gene results in a more than 70,000-fold reduction in luciferase expression in tobacco protoplasts. RNA probes carrying the sequence GCGC in place of the central four contiguous uridines of the 8-nucleotide U-rich motif fail to bind host protein at detectable levels, and the same mutation, when introduced into the PVX reporter virus, eliminates viral multiplication. Mutations of 1 or 2 nucleotides within the same four uridines reduced both binding of host proteins and replication of reporter virus. These results indicate that the 8-nucleotide U-rich motif within the PVX 3' UTR is important for some aspect of viral multiplication and suggest that host protein binding plays a role in the process.     

Origin and phylogeny of Oryza species with the CD genome based on multiple-gene sequence data

The CD genome species in the genus Oryza are endemic to Latin America, including O. alta, O. grandiglumis and O. latifolia. Origins and phylogenetic relationship of these species have long been in dispute and are still ambiguous due to their homogeneous genome type, similar morphological characteristics and overlapping distribution. In the present study, we sequenced two chloroplast fragments (matK and trnL-trnF) and portions of three nuclear genes (Adh1, Adh2 and GPA1) from sixteen accessions representing seven species with the C, CD, and E genomes, as well as one G genome species as the outgroup. Phylogenetic analyses using parsimony and distance methods strongly supported that the CD genome originated from a single hybridization event, and that the C genome species (O. officinalis or O. rhizomatis instead of O. eichingeri) served as the maternal parent while the E genome species (O. australiensis) was the paternal donor during the formation of CD genome. In addition, the consistent phylogenetic relationships among the CCDD species indicated that significant divergence existed between O. latifolia and the other two (O. alta and O. grandiglumis), which corroborated the suggestion of treating the latter two as a single species or as taxa within species.We thank Tao Sang of Michigan State University (East Lansing, USA) and Bao-rong Lu of Fudan University (Shanghai, China) for their encouragement and assistance. We are also grateful to the International Rice Research Institute (Manila, Philippines) for providing plant material for this study. This research was supported by the Chinese Academy of Sciences (kscxz-sw-101A), the National Natural Science Foundation of China (30025005) and the Program for Key International S & T Cooperation Project of P. R. China (2001CB711103).     

cGMP、cAMP及两种cAMP衍生物对大鼠肝细胞核体外转录系统的影响

 本文介绍了以α-鹅膏蕈碱和低浓度KCl为手段建立了RNA聚合酶Ⅰ、RNA聚合酶Ⅱ活性的细胞核转录系统进而研究了cGMP、cAMP、cAMP丁酯及cAMP硫代环磷酰二乙胺对大鼠肝细胞核中RNA聚合酶Ⅰ与Ⅱ活性的影响。结果显示cGMP可以提高RNA聚合酶Ⅰ的活性;cAMP主要提高RNA聚合酶Ⅱ的活极,而cAMP分子结构变化产生的丁酯及硫代环磷酰二乙胺衍生物可增强cAMP的这种作用,为深入研究cAMP的构效关系提供了实验依据。     

带γδ和αβ受体的T细胞（TCR）在人胎儿组织内分布的研究

本文用细胞免疫化学方法,在冰冻切片上,检测了胎儿不同组织和器官内带γδ和αβ受体的Ｔ细胞（ＴＣＲ）的分布,结果发现ＴＣＲ细胞的分布与,般Ｔ细胞不同,有相对固定的分布区,如胸腺内ＴＣＲ细胞主要分布在皮筋质交界处和髓质部;脾脏内的γδＴ主要位于边缘区,而αβＴ主要位于动脉周围淋巴鞘,在红髓和血窦两种细胞共存;淋巴结内只有少数ＴＣＲ细胞位于滤泡间或副皮质,滤泡内则未见。消化管内的ＴＣＲ细胞主要分布在小肠的固有膜,而胃、大肠和阑尾的固有膜内很少见;肝内ＴＣＲ细胞主要集中在血管和血窦周围;皮肤切片内的少数ＴＣＲ细胞见于真皮内,表皮基底层细胞内未见。这些细胞在胎儿期的免疫皮应及其生理功能还不清楚。     

γ—亚麻酸生产菌原生质体的制备及诱变条件的研究

将γ─亚麻酸生产菌Ｃｕｎｎｉｎｇｈａｍｅｌｌａｓｐｐ．８７６１进行原生质体化,并用物理方法进行诱变,使其在形态上产生了变异,γ─亚麻酸含量有所增加。本文报道了该菌株原生质体的制备,再生条件,以及诱变后形态变化,发酵性能。     

新疆罂粟属高山罂粟组分类研究

我国有罂粟属植物12种,新疆有10种,其中6种属于高山罂粟组。对新疆高山罂粟组植物的花葶进行了解剖研究,并用扫描电镜观察了花粉形态,结果表明,花葶中维管束的数量及排列方式在各种间存在着差异,可以作为一个鉴定特征。花粉上的小刺密度在放大12000倍时,在有种间存在着明显差异,可分类提供微观佐证。     

Development of a fluorescent probe hydrolysis-insulated isothermal PCR for rapid and sensitive on-site detection of African swine fever virus

Highlights
1. A probe-based insulated isothermal PCR (iiPCR) assay was developed for rapid and onsite detection of ASFV.
2. The developed iiPCR showed similar sensitivity and specificity with OIE recommended real-time PCR.
3. Blood samples could be directly applied as PCR template in iiPCR without DNA extraction.     

Superoxide anion burst and taxol production induced by Ce in suspension cultures of Taxus cuspidata

Generation of reactive oxygen species (ROS) induced by Ce⁴⁺ in suspension cultures of Taxus cuspidata was investigated. The burst of superoxide anions (O₂√⁻) occurred rapidly after the addition of Ce⁴⁺ and reached maximum at 4.3 h, while the total level of the cellular reactive oxygen species maintained unchanged. The intracellular superoxide dismutase (SOD) and catalase (CAT) were activated while the intra/extracellular peroxidases (PODs) were inhibited accompanying the O₂√⁻ burst. The pretreatment of the suspension cultures with diphenylene iodonium (DPI), a suicide inhibitor of the NADPH oxidase, blocked the O₂√⁻ burst, inhibiting the cell apoptosis and taxol production induced by Ce⁴⁺. These results show that NADPH oxidase played a key role in O₂√⁻ burst and O₂√⁻ served as a mediator of Ce⁴⁺ for cell apoptosis and taxol production. The pretreatments of the suspension cultures with anthracene-9-carboxylate, an ion-channel blocker, nifedipine, a Ca²⁺-channel blocker, neomycin, a phospholipase C (PLC) inhibitor, or suramin, a G-protein inhibitor, decreased O₂√⁻ burst induced by Ce⁴⁺. It is thus inferred that Ce⁴⁺-induced O₂√⁻ burst, which mediated cell apoptosis and taxol production by activating the ion-channels, PLC, G-proteins and NADPH oxidase.     

The intrinsic mechanisms underlying the maturation of programming sequential spikes at cerebellar Purkinje cells

Cerebellum is involved in the motion coordination and working memory, to which the programming of sequential spikes at Purkinje cells is essential. It is not clear about the intrinsic mechanisms underlying spike capacity and timing precision as well as their postnatal maturation. We investigated the programming and intrinsic property of sequential spikes at Purkinje neurons during postnatal development by whole-cell recording in cerebellar slices. Cerebellar Purkinje neurons demonstrate the increasing of spike capacity and timing precision, as well as the lowering of refractory periods and threshold potentials during the postnatal maturation. In addition, the correlation between spike parameters and intrinsic properties converts to be more linear. This postnatal plasticity of neuronal intrinsic properties improves the timing precision and capacity of spike programming at cerebellar Purkinje neurons.