破伤风毒素C基因的克隆及重组C蛋白的表达和免疫原性研究

对破伤风毒素C基因(TTC)进行了克隆,在大肠杆菌中构建了C基因-硫氧还蛋白融合表达系统并对其进行了重组表达,对表达后蛋白进行了纯化和rTTC的免疫原性初步研究。结果表明,该系统可高水平表达可溶性rTTC,rTTC表达量占可溶性蛋白的28.19%,纯化后纯度为96.92%;所获得的rTTC具有良好的免疫原性,可为开发新一代破伤风亚单位疫苗奠定基础。     

Bradyrhizobium japonicum基因工程菌株HN32中外源片段的来源检验

采用Southern转移,放射性同位素分子杂交等研究手段,对本课题组构建的一株高效固氮大豆根瘤菌HN32中的外源片段的来源进行了分析。研究结果证实:该外源片段确系来源于Sinorhizobium fredii B52菌株,并且在受体Bradyrhizobium japonicum 22—10菌株中本不存在。     

The Minimal Active Fragment of the CrylAi Toxin is Located Between 36 and 605

The novel crylAi gene that cloned from Bacillus thuringiensis strain SC6H8 encoded a protein exhibiting strong toxicity against Plutella xylostella and Chilo suppressalis in our previous study. Using the available information for the active fragments of other Cry toxins, eight truncated fragments were constructed to identify the minimal active fragment of CrylAi. All truncated fragments were expressed in Escherichia coli strain BL21 （DE3）, and the insecticidal activity against 2ndinstar P. xylostella larvae was assessed using full-length CrylAi as a positive control. The results indicate that the minimal active fragment of the CrylAi toxin against P. xylostella is located between amino acid residues 36^1 and 605^1, which is smaller than the regions previously reported for CrylA. The first two amino acids （34T and 35P） on helix a-1 and whole helix a-2 of domain I and sheet 13-32 of domain III are necessary for CrylAi toxin to keep its toxicity against P. xylostella.     

Distribution and biodiversity of soybean rhizobia in the soils of Shennongjia forest reserve,China

Soil samples were collected at an altitude of 500, 1,060, 1,500, 1,950, 2,400 and 3,100 m, respectively, from Shennongjia, a forest reserve in Hubei province (central China). Their corresponding pHs were 5.50, 4.91, 5.64, 5.28, 5.49 and 4.60. By using a plant trap method, a total of 25 soybean rhizobia were isolated from the soil above an altitude of 1,500 m and all identified to be Sinorhizobium fredii. Their genetic biodiversity was characterized by 16S–23S rDNA internally transcribed spacer (ITS) region polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and random amplification DNA (RAPD) analysis. All the tested strains produced a 2.1 kb 16S–23S rDNA ITS fragment. After digestion with three restriction endonucleases (HaeIII, MspI and CfoI), respectively, great variations in 16S–23S rDNA ITS PCR-RFLP patterns were observed. The tested strains could be differentiated into 11 ITS genotypes. The genotypes of rhizobia were not related to geographical location. Twelve primers were applied to RAPD analysis and a dendrogram was obtained, showing that all the strains (including reference strain S. fredii USDA205) were divided into two diverging groups. Moreover, each group could be further divided into two subgroups. Both RAPD and 16S–23S rDNA ITS PCR-RFLP analysis indicated that a high degree of genetic diversity existed among S. fredii strains isolated from Shennongjia virgin soils. Since Shennongjia is an unexploited forest region in central China and the gene centre of soybean is located in China, the symbiotic genes harboured by these strains may be of great importance and the rich diversity of these strains might contribute to the adaptation of soybean to an alpine environment.     

Production and analysis of reciprocal hybrids between <Emphasis Type="Italic">Asparagus officinalis</Emphasis> L. and <Emphasis Type="Italic">A. schoberioides</Emphasis> Kunth

The genus Asparagus (Asparagaceae) encompasses 100–300 species, including the important vegetable crop, A. officinalis (garden asparagus). Previous attempts to hybridize garden asparagus with A. densiflorus (Kunth) Jessop, with the aim of introducing disease resistance, were unsuccessful because of the failure of endosperm development. In the present study, reciprocal interspecific hybrids between garden asparagus and A. schoberioides Kunth were generated by hand pollination. The F1 hybrids were analyzed by using both morphological and molecular techniques. This is the first report describing the production of an interspecific hybrid between garden asparagus at the diploid level (2n = 2x = 20) and its diploid (2n = 2x = 20) wild relative, A. schoberioides. Morphological characteristics of candidate hybrids were a mixture of those found in the parents, and cytological and RFLP analyses confirmed that morphologically intermediate plants were indeed diploid hybrids of those two species. In other words, post-zygotic isolation is not complete between phylogenetically distinct these two species. Our results suggest that other Asparagus species may be capable of hybridizing with A. officinalis and that introducing wild characters through interspecific hybridization may offer advantages for breeding for novel traits. Takuro Ito and Toshinori Ochiai authors are contributed equally to this work     

.梅 花 品 种 资 源 核 心 种 质 构 建

梅花是中国观赏树木类品种资源研究较深入全面的种类之一，为了在有限的时间、财力的情况下，加强梅花品种资源的保护力度、提高利用效率，该研究利用梅花295个AFLP多态位点信息，采用系统取样的C和G取样策略，以Nei’s（1972）遗传距离进行的非加权成对算术平均法（UPGMA）聚类分析取样，从178个梅花品种及近缘（品）种构成的群体中，选取了52个品种样品作为梅花核心种质，总体取样比例为29.21%.遗传多样性和实用性检验分析结果表明:核心种质保留了梅花全部3个种系、5类18个品种类型；Nei’s (1973)基因多样度、Shannon’s信息指数、Simpson指数保留率分别为:99.90%、99.47%、99.99%.差异显著性检验结果显示核心种质与原全部种质的各项遗传参数差异不显著，表明该核心种质样品较好地保留了全部种质的遗传多样性和形态性状类型，为梅花等观赏植物改良提供了有益参考.      

Inheritance of Powdery Mildew Resistance in Cucumber （Cucumis sativus L.） and Development of an AFLP Marker for Resistance Detection

Cucumber powdery mildew is one of the most destructive diseases of cucumber throughout the world. In the present study, inheritance of powdery mildew resistance in three crosses, and linkage of resistance with amplified fragment length polymorphism （AFLP） markers are studied to formulate efficient strategies for breeding cultivars resistant to powdery mildew. The joint analysis of multiple generations and AFLP technique has been applied in this study. The best model is the one with two major genes, additive, dominant, and epistatic effects, plus polygenes with additive, dominant, and epistatic effects （E-l-0 model）. The heritabilities of the major genes varied from 64.26% to 97.82%, and susceptibility was incompletely dominant for the two major genes in the three crosses studied. The additive effects of the two major genes and the dominant effect of the second major gene were high, and the epistatic effect of the additive-dominant between the two major genes was the highest in cross I . In cross II, the absolute value of the additive effect, dominant effect, and potential ratio of the first major gene were far higher than those of the second major gene, and the epistatic effect of the additive-additive was the highest. The genetic parameters of the two major genes in cross III were similar to those in cross II. Correlation and regression analyses showed that marker E25/M63-103 was linked to a susceptible gene controlling powdery mildew resistance. The marker could account for 19.98% of the phenotypic variation. When the marker was tested on a diverse set of 29 cucumber lines, the correlation between phenotype and genotype was not significant, which suggested cultivar specialty of gene expression or different methods of resistance to powdery mildew. The target DNA fragment was 103 bp in length, and only a small part was found to be homologous to DNA in the other species evaluated, which indicated that it was unique to the cucumber genome.     

部分太湖流域粳稻品种的AFLP分析

 研究了24份太湖流域粳稻品种的AFLP，根据AFLP资料分析了太湖流域粳稻地方品种在推广品种中的渗透情况，结果表明大部分江、浙推广品种与地方品种间存在着相对较大的遗传距离，即地方品种在推广品种中的遗传组分较小，相反渗透进了较多的其他种质的遗传组分,这与推广品种的系谱分析结论一致。     

水稻抗白背飞虱基因Wbph2的初步定位

 鉴定了ARC10239/明恢63 F2群体对白背飞虱的抗性，混合F2感虫稻株的DNA，构建感虫池。应用129个RFLP探针，结合4种限制性内切酶，检测抗虫亲本ARC10239、感虫亲本明恢63和感虫DNA池的多态性。检测到6个阳性探针，分布在5个染色体上(染色体3、6、8、11和12)。应用阳性探针分析了F2群体的142个个体，发现抗虫基因Wbph2与第6染色体上的标记RZ667、RG64和RG264连锁,其中， Wbph2与RZ667的遗传距离为25.6 cM (LOD=4.50)。     

Temporal and seasonal change in microbial community structure of an undisturbed,disturbed, and carbon-amended pasture soil

Disturbance and change to C inputs can alter microbial community structure and impact ecosystem function. Particularly in temperate regions, seasonal change also has an effect on microbial communities both directly through climate and indirectly through plant function. The temporal change in microbial communities of an undisturbed pasture, disturbed pasture (similar to a single tillage event) and pasture soil amended with two forms of particulate carbon were monitored over eight consecutive seasons after grass was reestablished. The soil microbial community was assessed by a DNA fingerprinting technique (terminal restriction fragment length polymorphism, TRFLP) of bacterial, fungal and archaeal communities, and also from phospholipid fatty acid (PLFA) analysis. The single disturbance had a significant effect on fungal microbial community structure (by TRFLP) and significantly decreased the fungal:bacterial ratio. Though the change was relatively small, it persisted throughout the sampling period. Nitrate was also higher on the disturbed treatment providing evidence for the theory that changes to fungal:bacterial ratios can alter nutrient cycling and retention. Fungal communities were the most altered by the C amendments, and while bacteria were also affected by the C amendments, seasonal change was a greater cause of variation. Correlation to soil and climatic variables explained more of the total variability for PLFA (78% for all treatments) than bacterial (50%), fungal (35%) and archaeal (14%) restriction fragments. Most climate and soil variables explained significant variation for seasonal patterns in the multivariate community structures but measurements of soil moisture were important for all communities while pH was relatively more important for bacteria, temperature for fungi, and soil C:N ratio for archaea. Autumn was particularly distinct from other seasons for bacteria (less so for the fungal community) and although there was seasonal change in pH suggesting pasture management was a factor, the significant correlation of other soil characteristics suggests that plant physiological changes (most probably root exudates) also played a significant role. The large change in the saprotrophic fungal community due to the particulate C addition but minor seasonal change would tend to suggest that the fungal community may be more responsive to changes in litter inputs rather than root exudates while the reverse is true for bacteria.