黄连中原小檗碱型生物碱分子构型和结构研究

本文以中药黄连中原小檗碱型生物碱为研究对象,采用重结晶的方法,获得盐酸小檗碱和盐酸药根碱的针状晶体.应用量子力学计算研究盐酸小檗碱和盐酸药根碱的分子构型,同时使用粉末X射线衍射研究结晶体的初步晶体构型.实验表明两种晶体同属三斜晶系,盐酸药根碱的晶胞参数为:a＝15.417 ,b＝14.850 ,c＝8.013 ,α＝114.746°,β＝94.436°,γ＝95.071°;盐酸小檗碱的晶胞参数为:a＝13.608 ,b＝11.080 ,c＝12.775 ,α＝80.360°,β＝105.110°,γ＝111.640°.     

Selection and identification of a new adhesion protein of Cryptosporidium parvum from a cDNA library by ribosome display

In this study, we described a novel display method to identify surface adhesion proteins of Cryptosporidium parvum. A cDNA library of the sporozoite and oocyst stages of C. parvum was expressed on ribosome and selectively and specifically screened with intestinal epithelial cells (IECs) from newborn Cryptosporidium-free Holstein calves. Proteins were then enriched using a multi-step panning procedure. A new surface adherence protein of C. parvum was selected, named Cp20. Sequence analyses showed that Cp20 has a N-terminal signal peptide and four transmembrane regions. Indirect immunofluorescence assay (IFA) using an antibody specific for rCp20 demonstrated that the antibody specifically bound to the surface of sporozoites and oocysts. The recombinant plasmid pVAX1-Cp20 was constructed to examine the potential of the Cp20 gene as a target for specific preventive and therapeutic measures for cryptosporidiosis. The in vivo efficacies of the DNA vaccine was tested in BALB/c mice. The results indicated that the DNA vaccine elicited significant antibody responses and specific cellular responses when compared to control mice that received vector only or PBS. The DNA vaccine induced strong protective immune response against C. parvum and lower level of the oocysts shedding after challenge infection. This study suggested that Cp20 could serve as an effective target for specific preventive and therapeutic measures for cryptosporidiosis.     

产喜树碱喜树内生真菌的筛选及喜树内生真菌的SRAP分析

生活在宿主植物里的内生真菌是很重要的药用资源。喜树是中国的传统药用植物。从喜树植物中分离得到了大约50种菌株,其中一株产喜树碱的菌株通过形态学鉴定为青霉属,这是首次在喜树植物中发现产喜树碱的青霉属菌株。为研究简单序列重复相关序列扩增多态性(SRAP)方法在喜树内生真菌中应用的可行性,选择了十株喜树内生真菌进行SRAP多态性分析。SRAP引物共扩增出1 295条带,而这些菌株也被分为三大类。这些结果表明,SRAP研究喜树内生真菌具有高效性,是讨论喜树内生真菌的遗传多样性的有效方法。     

THE PEST STATUS OF BEMISIA TABACI IN CHINA AND NON‐CHEMICAL CONTROL STRATEGIES*

Abstract Bemisia tabaci (Gennadius) has been considered as a serious pest in all of tropical, subtropical and temperate regions of the world. B. tabaci first recorded as early as in 1940s in China and has been reported as a pest of various crops in 22 provinces or cities. But only recently it has become a severe problem for vegetable and ornamental crops in Guangdong and Beijing. In China B. tabaci is known to transmit at least 5 plant viruses, including tomato yellow leaf curl virus (TYLCV), tomato leaf curl virus (TomLCV), squash leaf curl virus (SqLCV‐C). So far, approximately 18 parasitoids, 17 predators and 1 pathogenic fungus were recorded in China. This paper presents an overview of B. tabaci as a pest and virus vector in China, with special attention given to non‐chemical control strategies.     

康氏木霉纤维素酶系中C_x酶多分子型的分离纯化及某些性质

从康氐木霉(Trichoderma k(?)ningii)白色变异株As 3.4001的粗酶制剂中,获得了纤维素酶系中的一组C_x酶(C_(x1) C_(x2) C_(x3) C_(x4))。分离步骤包括Sephadex G-75凝胶过滤,DEAESephadex A-50离子交换层析,ConA-Sepharose亲合层析,SE-Sephadex C-50离子交换层析及聚丙烯酰胺凝胶电泳。C_(x1)与C_(x2)的分子量不同而所带电荷相同,它们的分子量各自为44,500和34,000。C_(x2)—C_(x4)的分子量相同而所带电荷不同。纯化的C_(x1)—C_(x4)经聚丙烯酰胺凝胶电泳鉴定为单带。比较它们对羧甲基纤维素钠(CMC-Na)的糖化力及液化力表明在作用方式的随机性上C_(x2)>C_(x3)>C_(x1)>C_(x4)。     

杀虫剂抗性: 遗传学、基因组学及应用启示

杀虫剂抗性已成为害虫防治工作需要解决的一个重要问题,也是一种人为的、自然选择的重要的进化现象,开展抗药性的研究不仅为抗性的监测、治理和农药工业的发展提供科学参考,还可以揭示生物进化的一些基本规律。在过去的10年,昆虫对许多化学杀虫剂抗药性的分子基础得到了进一步阐明,已从果蝇Drosophila melanogaster中克隆了杀虫剂的靶标基因,还查明了一些害虫的与抗性相关联的基因突变。最近,随着经注释的昆虫基因组的出现,由复杂多基因酶系如酯酶、细胞色素P450酶及谷胱甘肽S-转移酶介导的抗性的机制有了突破性的进展,有关杀虫剂抗性的进化以及抗性基因的传播模式也逐步得到揭示。基因组技术在揭示昆虫其他可能的抗药性机制以及在发现新的杀虫剂靶标方面将发挥更大的作用。     

Developing DNA barcodes for species identification in Podophylloideae (Berberidaceae)

Species of Podophyllum, Dysosma, Sinopodophyllum, and Diphylleia, genera from Podophylloideae of Berberidaceae, have long been used in traditional herbal medicine in East Asia and/or North America. Accurate identification of the species of these four genera is crucial to their medicinal uses. In this study, we tested the utility of nine barcodes (matK, rbcL, atpH-atpI, rpl32-trnL^UAG, rps18-clpp, trnL-trnF, trnL-ndhJ, trnS-trnfM, and internal transcribed spacer (ITS)) to discriminate different species of Podophylloideae. Thirty-six individuals representing 12 species of Podophylloideae were collected from different locations in China, Japan, and North America. We assessed the feasibility of amplification and sequencing of all markers, examined the levels of the barcoding gap based on DNA sequence divergence between ranges of intra- and interspecific variation using pairwise distances, and further evaluated successful identifications using each barcode by similarity-based and tree-based methods. Results showed that nine barcodes, except rps18-clpp, have a high level of primer universality and sequencing success. As a single barcode, ITS has the most variable sites, greater intra- and interspecific divergences, and the highest species discrimination rate (83%), followed by matKwhich has moderate variation and also high species discrimination rates. However, these species can also be discriminated by ITS alone, except Dysosma versipellis (Hance) M. Cheng ex T. S. Ying and D. pleiantha (Hance) Woodson. The combination of ITS + matK did not improve species resolution over ITS alone. Thus, we propose that ITS may be used as a sole region for identification of most species in Podophylloideae. The failure of ITS to distinguish D. versipellis and D. pleiantha is likely attributed to incomplete lineage sorting due to recent divergence of the two species.     

3个中山杉杂种无性系及其亲本的光合特性比较

对‘中山杉405’(Taxodium‘Zhongshanshan 405’)、‘中山杉406’(T.‘Zhongshanshan 406’)和‘中山杉407’(T.‘Zhongshanshan 407’)及其母本墨西哥落羽杉(T.mucronatum Tenore)和父本落羽杉〔T.distichum(L.)Rich.〕在7月份、9月份和10月份的光合参数进行了测定,并比较了3个无性系净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)和水分利用效率(WUE)的杂种优势。结果显示:不同月份3个无性系及其亲本的Pn值有明显差异,导致光响应曲线明显不同,但3个无性系的光响应曲线基本一致;3个无性系的Pn值在不同月份都高于落羽杉,且‘中山杉405’的Pn值高于另2个无性系。3个无性系及其亲本的光补偿点总体上表现为7月份最高、10月份最低,其中7月份墨西哥落羽杉的光补偿点显著高于3个无性系及落羽杉(P<0.05);3个无性系和墨西哥落羽杉的光饱和点则为9月份最高、7月份最低,而落羽杉的光饱和点为9月份最高、10月份最低。7月份3个无性系与其亲本的Pn日变化曲线均呈不规则型,低谷出现在中午12:00左右;9月份和10月份3个无性系和墨西哥落羽杉的Pn日变化曲线均呈单峰型,峰值出现在上午10:00;落羽杉的Pn值从上午8:00开始呈持续降低的趋势,且全天都明显低于其他树种。不同月份3个无性系的Pn、Gs、Tr和WUE的杂种优势有明显差异,总体上看,‘中山杉405’的Pn、Gs和Tr在3个月份中均保持较高的杂种优势率,而‘中山杉406’9月份的杂种优势率最高,‘中山杉407’则在7月份杂种优势率最高;3个无性系的WUE仅在10月份有明显的杂种优势,且‘中山杉405’和‘中山杉406’的杂种优势率远高于‘中山杉407’;‘中山杉405’各项指标的杂种优势率总体上高于另2个无性系。研究结果表明:3个中山杉无性系的光合能力接近其母本墨西哥落羽杉、高于父本落羽杉,其中‘中山杉405’杂种优势明显。     

白蚁的分子生物学研究进展

白蚁是危险性社会昆虫,建立安全有效的白蚁防治方法有赖多学科的参与,分子生物学已经成为白蚁研究的重要工具。目前,DNA序列分析方法已应用于白蚁鉴定和分类,其中线粒体基因是最通用的分子标记;基因工程技术成功构建了可用于白蚁防治的白蚁肠道工程菌;Hexamerin、COX Ⅲ、纤维素酶等白蚁功能基因以及白蚁品级分化相关的若干蛋白相继得到了分离和鉴定。文章从白蚁分类、防治、功能基因、品级分化4个方面综述白蚁分子生物学的研究进展,为白蚁的防治提供新的方法和思路。