Transgenic grapevine plants expressing a rice chitinase with enhanced resistance to fungal pathogens

 The rice chitinase gene (RCC2), classified as class I chitinase, was introduced into the somatic embryos of grapevine (Vitis vinifera L. cv. Neo Muscut) by Agrobacterium infection. After co-cultivation with Agrobacterium, somatic embryos were transferred onto Murashige and Skoog hormone-free medium supplemented with 50 mg/l kanamycin. Transformed secondary or tertiary embryos were selected, and then more than 20 transgenic plantlets were recovered. Two transformants showed enhanced resistance against powdery mildew caused by Uncinula necator. Few disease symptoms were observed on leaves of these transformants compared with those of the non-transformant, although browning and necrotic symptoms, which seemed to constitute a hypersensitive reaction, were observed. Scanning electron microscopic observation revealed that conidial germination, mycelial growth and conidial formation were suppressed on the leaf surface of the transformant. The transgenic grapevines obtained also exhibited slight resistance against Elisinoe ampelina inducing anthracnose, resulting in a reduction in disease lesions. The relationship between the expression of the foreign chitinase gene and the disease resistance is discussed. Received: 5 April 1999 / Revision received: 13 September 1999 / Accepted: 6 October 1999     

利用鸡胚浸出液制作细菌培养基的研究

利用鸡胚浸出液制作的肉汤和琼脂平板培养基均能适合副鸡嗜血杆菌、大肠杆菌和沙门氏菌的生长,用平板计数法、麦氏比浊法和离心称重法分别对副鸡嗜血杆菌、大肠杆菌和沙门氏菌的培养产物进行含菌量、菌泥湿重测定,结果表明,鸡胚浸出济培养基培养的沙门氏菌产量是普通培养基培养的沙门氏菌产量的20倍以上,培养的副鸡嗜血杆菌产量是普通培养的3倍以上,而鸡胚浸出液培养基与普通培养基对大肠杆菌的培养数量则无明显差异。     

Synthesis of chicken galanin and its N- and C-terminal segments,and preparation of their antisera

Summary We describe the synthesis of the first avian galanin (GAL), chicken GAL, and its N-terminal and C-terminal segments by solid-phase synthesis, using Boc/Bzl amino acid protection groups and MBHA resin. The three peptides were prepared with purities of over 97%, as determined by RP-HPLC, HPCE, FAB-MS or ESI-MS and amino acid analysis. Antibodies against these synthetic peptides were raised in rabbits and used for immunohistochemical localization of GAL-immunoreactive neurons in chicken brain.     

一种适于植物幼胚mRNA整体原位杂交的方法

A mRNA whole-mount in situ hybridization method is reported here for quick, direct analysis of the spatial and temporal mRNA expression patterns in plant young embryos. A cDNA clone THE3 (tobacco heart embryo 3) was isolated by differential screening from tobacco (Nicotiana tabacum L.) heart embryo cDNA library as compared with the globular embryo cDNA library. The distribution of THE3 mRNA in tobacco heart embryos and globular embryos was investigated by a whole-mount in situ hybridization technique, showing that THE 3 is preferentially expressed in heart embryos.     

我国部分地方鸡种肤色伴性遗传初步观察

对泰和鸡、仙居鸡、固始鸡、北京油鸡、萧山鸡、狼山鸡（Ｎ系）进行肤色伴性遗传观察,结果表明：泰和鸡常染色体上含有１对黑色素基因ＰＰ,仙居鸡、萧山鸡、北京油鸡性染色体上含有Ｉｄ抑制色素基因,狼山鸡、固始鸡、泰和鸡性染色体上含有ｉｄ基因,泰和鸡♂与含有Ｉｄ基因鸡种（仙居鸡、萧山鸡、北京油鸡）♀杂交,Ｆ１代肤色能自别雌雄,公鸡为黄皮肤,母鸡为黑皮肤。     

藏鸡群体遗传多样性研究

藏鸡的体型外貌和生活习性与红色原鸡非常相似,是具有自己独特群体遗传特性的高原地方鸡种。为了有效保护并合理利用这一遗传资源,我们采用多重PCR与半自动荧光标记微卫星聚丙烯酰胺凝胶电泳相结合的方法检测了20个微卫星基因座的多态性,并随机抽取藏鸡群体中部分个体进行个体体形特征与生产性能的统计。结果表明藏鸡群体的20个微卫星基因座的多态等位基因数为4~10个,平均值为7.25个/基因座,多态信息含量（CPI）和杂合度（H）平均值分别为0.67、0.74。大染色体较小染色体的微卫星标记多态性程度要高。藏鸡群体的微卫星基因座多态性丰富,也解释了生产性能不均,外貌表现迥异的群体遗传特性。 Abstract: Morphological traits and living habit of Tibetan chicken, which is an aboriginal chicken breed on plateau with its own characteristic populational genetic features, are in great common with the Red Jungle Fowl, the assumed ancestry of domestic chicken. To fully exploit this chicken resource, Multiplex PCR with semi-automated polyacrilamide gel electrophoresis (PAGE) using fluorescently labeled microsatellite primers was used to detect the polymorphism at 20 microsatellite loci. At the same time, we randomly test the individual morphology and performance. It showed that numbers of polymorphic alleles were 4-10, with mean value 7.25 per locus. Polymorphism Information Content (CPI) and Heterozygosity (H) had mean values 0.67 and 0.74, respectively. Macrochromosomes had relatively higher polymorphism than microchromosomes(P>0.05). In all, high polymorphisms at microsatellite loci related to the uneven production performance and morphological discrepancy of population genetic characteristics in Tibetan chicken.     

家蚕母性基因的表达序列标签分析

为了研究家蚕Bombyx mori卵中储存的母性遗传信息,以未受精的肾脏型和正常型蚕卵为材料构建了cDNA文库,并对其随机测序,共获得391条表达序列标签(EST)序列,经拼接后得到374条非重复序列。生物信息学分析发现,172个非重复序列与国际数据库GenBank和silkbase中的已知基因具有较高的相似性。除3个已知母性基因外,其余主要与DNA复制、能量代谢、信号转导、转录、蛋白质结构以及胚胎发育中个体形成等相关。该结果可为进一步研究家蚕胚胎早期发育的调控提供基础数据。     

涕灭威及其复合污染对斑马鱼胚胎DNA的影响

研究了一种毒性很高的氨基甲酸酯类杀虫剂--涕灭威和广泛使用的阴离子表面活性剂十二烷基苯磺酸钠(SDBS)组成的复合污染体系对斑马鱼胚胎DNA的影响．结果表明,涕灭威对斑马鱼胚胎DNA的损伤随浓度增大而加重,但低浓度涕灭威在短时间内造成的DNA单链断裂是可以修复的,高浓度则导致难以修复的双链断裂．一定浓度(20mg·L^-1)的SDBS在复合污染体系中能减弱涕灭威的毒性．     

氯化钠密度梯度离心法制备用于显微注射的外源DNA片段

DNA显微注射是生产转基因动物最可靠和最常使用的一种方法,外源DNA的纯度对显微注射的成功起着至关重要的作用。本文介绍用氯化钠密度梯度离心的方法制备用于显微注射的外源DNA片段。与传统的琼脂糖凝胶回收的方法相比较,用此方法制备的外源DNA片段对小鼠受精卵进行显微注射后,受卵体母鼠的胚胎存活率,以及子代小鼠的外源基因整合率均有明显的提高。这一方法可为进一步提高转基因动物的成功率,提供方法学上的参考。 Abstract:DNA microinjection is the most popular and reliable method of producing transgenic animals.The purity of foreign DNA plays an important role for the success of microinjection.In this study,we introduced the use of sodium chloride step gradients in fractionating foreign DNA fragment for microinjection.The data demonstrated that,compared with the conventional agarose gel extraction method,NaCl purification scheme of toreign DNA could improve the treated embryo survival and foreign DNA intergration rate markedly.     

鸡粪沼气池产甲烷菌多样性

采用分子生物学方法构建16SrDNA基因文库,研究鸡粪沼气池发酵液中产甲烷菌的菌群结构。随机分析文库中50个克隆的16SrDNA基因序列,结果发现,其中46个克隆属于产甲烷菌属,与Methanogenium marinum strain AK-1菌株的相似性为99%-100%,占总数的92%;3个克隆(KD525、KD526和KD567)属于甲烷袋状菌属,与Methanoculleus sp.dm2菌株的相似性均为99%,占总数的6%;1个克隆(KD519)属于甲烷粒菌属,与Methanocorpusculum bavaricum菌株的相似性为99%,占总数的2%。另外,同样分析了样品中甲基辅酶M还原酶alpha亚基mcrA基因氨基酸序列的差异。气相色谱法分析结果显示,发酵液中甲酸含量为28.85g/L,约占总有机酸含量的81.7%;沼气的主要成分为甲烷、二氧化碳和氢气,分别约占总气体量的55.5%、41.1%和3.2%。