Engineering of a Single Chain Variable Fragment Antibody Specific for the Citrus tristeza virus and its Expression in Escherichia coli and Nicotiana tabacum

Citrus tristeza virus (CTV) is one of the most destructive citrus virus diseases in the world. The construction of an engineered antibody, EMBL accession number AJ278109, able to specifically recognize its antigen, i.e. the coat protein of CTV, directly on infected plant material without any purification or manipulation of the entire woody plant. The potential uses of this engineered antibody are discussed.     

Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .     

Bt对小地考虑呼吸作用的影响

利用红外CO2分析仪测定小地老虎幼虫呼吸节律的变化结果显示：小地老虎幼虫呼吸节律属于不规则波动型，取食含Bt烟叶的幼虫呼出的CO2量显著低于对照组，且呼吸节律的波动性也明显增大，说明小地考虑幼虫取食Bt后，呼吸代谢受到一定干扰，出现呼吸水平降低及代谢紊乱现象。     

梨自交不亲和强度不同品种花柱S糖蛋白含量的差异

 提取梨( Pyrus) 自交不亲和强度不同品种花柱可溶性蛋白质, 应用等电聚焦电泳法在凝胶板上分离出S 糖蛋白, 用图像解析法进行定量。结果表明, 花柱总S 糖蛋白含量因品种而异, 自交不亲和强、弱及自交亲和的品种每1 􀀁g 花柱可溶性蛋白中含总S 糖蛋白分别为􀀂1. 3 ng 、0. 7 ~ 0. 9 ng、 0. 7 ng 。不同的S 基因所表达的S 糖蛋白量差异显著, 即使是同一S 基因所对应的S 糖蛋白量也因品种而异。     

苹果梨中多酚氧化酶酶学特性的研究

 以苹果梨中的多酚氧化酶为研究对象, 对其酶的活性变化及其特性的研究表明, 苹果梨中的多酚氧化酶的最适pH 值是4. 6 , 最适温度是40 ℃; 亚硫酸氢钠、维生素C、柠檬酸对苹果梨中多酚氧化酶的抑制作用依次减弱, 硫酸钠几乎无抑制作用。不同底物和同底物不同浓度影响多酚氧化酶特性。     

核桃早实性状的RAPD分析

 用BSA (Bulked Segregant Analysis) 法获得了与核桃早实性状相关的RAPD 标记OPB208₉₀₀。混合分离群体的混合样来自早实品种辽1 的自然授粉实生后代。用220 个随机引物扩增早实和晚实混合样, 只有引物OPB208 ( 5. GTCCACACGG 3. ) 在早实混合样及其个体上扩增出了约900 bp 的特异带OPB208₉₀₀, 而在晚实混合样及其个体上无此特异带。用OPB208 为引物, 以19 个品种和3 个株系的DNA 为模板进行扩增,特异带OPB208₉₀₀在17 个早实品种中的12 个个体上出现, 而5 个晚实品种( 系) 上均无此带。     

Spatial dependence and the relationship of soil organic carbon and soil moisture in the Luquillo Experimental Forest,Puerto Rico

We used geo-spatial statistical techniques to examine the spatial variation and relationship of soil organic carbon (SOC) and soil moisture (SM) in the Luquillo Experimental Forest (LEF), Puerto Rico, in order to test the hypothesis that mountainous terrain introduces spatial autocorrelation and crosscorrelation in ecosystem and soil properties. Soil samples (n = 100) were collected from the LEF in the summer of 1998 and analyzed for SOC, SM, and bulk density (BD). A global positioning system was used to georeference the location of each sampling site. At each site, elevation, slope and aspect were recorded. We calculated the isotropic and anisotropic semivariograms of soil and topographic properties, as well as the cross-variograms between SOC and SM, and between SOC and elevation. Then we used four models (random, linear, spherical and wave/hole) to test the semi-variances of SOC, SM, BD, elevation, slope and aspect for spatial dependence. Our results indicate that all the studied properties except slope angle exhibit spatial dependence within the scale of sampling (200 – 1000 m sampling interval). The spatially structured variance (the variance due to the location of sampling sites) accounted for a large proportion of the sample variance for elevation (99%), BD (90%), SOC (68%), aspect (56%) and SM (44%). The ranges of spatial dependence (the distances within which parameters are spatially dependent) for aspect, SOC, elevation, SM, and BD were 9810 m, 3070 m, 1120 m, 930 m and 430 m, respectively. Cross correlograms indicate that SOC varies closely with elevation and SM depending on the distances between samples. The correlation can shift from positive to negative as the separation distance increases. Larger ranges of spatial dependence of SOC, aspect and elevation indicate that the distribution of SOC in the LEF is determined by a combination of biotic (e.g., litterfall) and abiotic factors (e.g., microclimate and topographic features) related to elevation and aspect. This demonstrates the importance of both elevation and topographic gradients in controlling climate, vegetation distribution and soil properties as well as the associated biogeochemical processes in the LEF.This revised version was published online in May 2005 with corrections to the Cover Date.     

Approach of allelopathy study with Arabidopsis thaliana (L.) Hevnh. and Neurospora crassa

Allelopathy in Arabidopsis seeds was investigated in the present study because there are few available reports of allelopathy studies regarding the early development stage of the plant life cycle, and because Arabidopsis is widely used as a model plant for molecular biology. When cockscomb ( Celosia cristata ) was incubated together with Arabidopsis seeds, the growth of cockscomb was promoted by Arabidopsis seeds. Exudate of Arabidopsis seeds showed a promotive activity on the growth of cockscomb and Arabidopsis itself. A new method for studying allelopathic mechanisms involving the interaction of the plant Arabidopsis thaliana (L.) Hevnh. and the fungus Neurospora crassa was suggested. Arabidopsis seed exudate and the water-soluble fraction of the exudate promoted Arabidopsis growth and reduced fungal germination, indicating that the seed exudate has a species-selective activity. Conversely, the ethylacetate-soluble fraction inhibited growth of all tested materials in the current study. These results demonstrate that the water-soluble substance(s) released from Arabidopsis seeds have species-selective activity on growth of both plants and microorganisms.     

外源绿原酸对苹果抗病相关酶的影响

采用离体培养方法对富士、红星和八棱海棠进行绿原酸处理,研究苹果在外源绿原酸处理下POD、PPO、PAL活性的动态变化以及绿原酸含量的变化。结果表明,富士以0.0001%绿原酸、红星以0.0008%绿原酸、八棱海棠以0.0004%绿原酸处理后,其PAL活性均显著提高,苹果体内也因此而提高了绿原酸水平。     

引起糖甜菜细菌性叶斑病的萎蔫短小杆菌新致病变种

 1995年在内蒙古临河市新发现了糖甜菜细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析、DNA G+C mol%和DNA-DNA同源性测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛、亚极生或侧生,结合其生理生化特性、细胞化学成分和DNA G+C mol%和DNA-DNA同源性测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur. flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下的一个新的致病变种,定名为Curtobacterium flaccumfaciens pv. beticola pv. nov. Chen et al.,2000(萎蔫短小杆菌糖甜菜致病变种)。