Analysis of genetic diversity of soybean germplasm from five different origins using RAPD markers

The pattern of genetic diversity among 92 genotypes of soybean from 5 different origins/sources (Pakistan, the USA, Asian Vegetable Research Development Centre (AVRDC), Japan and North Korea) was analyzed using randomly amplified polymorphic DNA markers. Out of 20 random primers 6 tested, 10 were polymorphic among genotypes and they yielded 107 markers, with an average of 10.7 markers per primer. The proportion of polymorphic bands within genotypes ranged from 0.47 to 0.71 with an average of 0.59. Pakistani and US genotypes exhibited the highest number of polymorphic bands (95%), while North Korean genotypes revealed the lowest (60%). The mean band frequency of the primers among genotypes was 0.57 with a range of 0.08–0.99. The Shannon’s index and Nei’s genetic diversity index revealed that primer OPF-06 showed maximum genetic diversity among the genotypes. Dendrogram constructed using Unweighted Pair Group Mean Average (UPGMA) method divided the genotypes into 5 main groups consisting of 13 clusters. The results of cluster analysis indicated that the genetic diversity between Pakistani and US or AVRDC genotypes is much larger than that between Pakistani and North Korean or Japanese genotypes. The Pakistani genotypes had distinct bands from plant introductions. Therefore, the Pakistani genotypes may be useful to soybean breeders.     

海岛型城市的土壤侵蚀状况及主要影响因子分析——以福建省平潭岛为例

为研究海岛型城市建设发展过程中的水土流失状况,将地理信息系统(GIS)与通用土壤流失方程(USLE)相结合,以福建省典型海岛——平潭岛作为研究区域,运用GIS建立平潭岛研究区的基础地理数据库,利用ArcGIS的栅格数据空间分析功能,对1990年与2013年的土壤侵蚀量进行了估算和分析。结果表明:平潭岛1990年平均土壤侵蚀模数为1 674.64t/(km~2·a),2013年平均土壤侵蚀模数为735t/(km~2·a),均属于轻度侵蚀。微度侵蚀从1990年的72.38%增加到2013年的79.31%。而在轻度侵蚀、中度侵蚀、强度侵蚀、极强度侵蚀和剧烈侵蚀方面均有所下降。1990年、2013年均是强度以上侵蚀区域对土壤侵蚀量的贡献最大。土地利用变化、降雨量和坡度变化对土壤侵蚀的作用显著。该研究可为平潭岛的经济开发建设、土地合理利用,以及为其他典型海岛在城市化进程中提出最佳的水土保持管理措施提供参考。     

菠菜中3种新烟碱类杀虫剂多残留分析方法研究

寄生隐丛赤壳菌是引起板栗疫病的致病菌。为建立该菌的分子检测技术,本研究首先采用通用引物 ITS1／ITS4对分离自四川雅安、泸州及重庆的寄生隐丛赤壳菌及其他参试菌株的 ITS 区进行 PCR 扩增和测序比对。根据该片段与 GenBank 中隐丛赤壳属其他种的 ITS 序列差异,设计了寄生隐丛赤壳菌的特异性引物 ITSP1／ITSP2,片段扩增大小为462 bp。利用该引物对菌株基因组 DNA 进行扩增,可以将寄生隐丛赤壳菌与其他参试菌区分开,检测灵敏度达30 pg。而以引物 ITS1／ITS4和 ITSP1／ITSP2进行的巢氏 PCR,可检测到30 fg 基因组 DNA,其灵敏度较常规 PCR 提高了1000倍。利用巢氏 PCR 检测体系对发病程度不同的组织和携菌组织进行检测,均能快速稳定地检测出寄生隐丛赤壳菌。     

Usefulness of single copy genes containing introns in Phytophthora for the development of detection tools for the regulated species P. ramorum and P. fragariae

Introns are generally highly polymorphic regions within genes and were proven to be of great interest for discriminating among phylogenetically-close Phytophthora species. Phytophthora ramorum and P. fragariae are considered as quarantine pathogens by the European Union and accurate detection tools are therefore necessary for their monitoring. From introns located in different single copy genes (GPA1, RAS-like, and TRP1), we developed a series of PCR primers specific to P. ramorum and P. fragariae. The specificity of these primers was successfully checked with a wide collection of Phytophthora isolates and a protocol was developed to detect both pathogens directly in infected plant tissues. These genes should be of particular interest for the development of additional species-specific detection tools within the Phytophthora genus.     

Rapid bidirectional allele-specific PCR identification for triazine resistance in higher plants

Triazine resistance is reported to be due to chloroplast herbicide target insensitivity in most species, and this is most often caused by a Ser(264)-Gly mutation at the D1 protein. In order to ascertain whether this mutation is really predominant amongst resistant plants, and also for gene flow studies, a rapid test is needed that allows the testing of large quantities of plants. Here a bidirectional allele-specific PCR (polymerase chain reaction) identification is proposed. The designed primers were shown to be universal in the three grass and three broadleaf species examined.     

PCR-based specific and sensitive detection of Pectobacterium carotovorum ssp. carotovorum by primers generated from a URP-PCR fingerprinting-derived polymorphic band

A 24-mer primer pair was generated by sequencing a URP-PCR fingerprinting-derived polymorphic band that is uniquely shared in Pectobacterium carotovorum ssp . carotovorum strains (Pcc). The primer set (EXPCCF/EXPCCR) amplified a single band of expected size (0·55 kb) from genomic DNA obtained from 29 Pcc strains and three Pectobacterium carotovorum ssp. wasabiae (Pcw) strains, but not from other P. carotovorum subspecies atrosepticum , betavasculorum or odoriferum , or from other Erwinia spp. or bacterial genera. The Rsa I digestion profile of the amplified bands divided Pcc strains into five groups with a unique profile from Pcw strains. First-round PCR detected between 5 × 10² and 1 × 10³ colony forming units (CFU) mL⁻¹ and detection sensitivity was increased to as few as 2–4 CFU mL⁻¹ after second-round (nested) PCR. This PCR protocol was used directly to detect Pcc strains in infected plant tissues.     

A novel pair of universal primers for the detection of potyviruses

A novel pair of universal primers was developed to detect potyvirus species after conserved sites were identified using all full‐length potyvirus sequences available by 2005. The breadth of specificity of the new primers, NIb2F and NIb3R, was investigated and compared with the specificity of two routinely used primer pairs in plant virus diagnostic laboratories. RNA from 40 potyvirus isolates representing 23 recognized and three possible new species was tested. Reactions with NIb2F and NIb3R produced amplicons of 350 bp from all 40 virus isolates tested. Reactions with the previously published WCIEN and Potyvirid primers amplified cDNA from 32 and 21 isolates, representing possibly 21 and 15 species, respectively. The identity of 12 unknown potyvirus isolates was confirmed by sequencing and three were found to be potentially distinct potyvirus species. Gel banding patterns from reactions with NIb2F and NIb3R were simpler to interpret than those from reactions with the other two primer sets; fewer products were visible and the cDNA fragments were less variable in size. RT‐PCR with the novel primers is predicted to be able to detect virus isolates from all major groups within the genus Potyvirus and its reliability makes it well suited for use as a routine diagnostic assay.     

Genotyping Cephalosporium gramineum and development of a marker for molecular diagnosis

This study investigated the genetic variation of 40 isolates of Cephalosporium gramineum, the causal agent of cephalosporium stripe disease of wheat, based on variations in internal transcribed spacers (ITS) and intergenic spacers (IGS) of rDNA. Of the isolates, 29 were from Japan and the rest from the USA and Europe. The ITS region was about 600 bp and almost identical among these isolates. In the IGS region (～5 kbp), restriction fragment length polymorphism analysis detected four genotypes among the 40 isolates. One representative isolate was selected from each of the four genotypes, and the IGS region was sequenced. Attempts to design a genotype‐specific marker based on the size of PCR products amplified with selected primers failed to differentiate among the four genotypes. Alternatively, a species‐specific primer set (CGIGS1 and CGIGS2) was developed that annealed within the conserved region, producing a DNA fragment of about 1·8 kbp. Tests of this primer set on a wide range of other fungi from 11 genera confirmed that it was specific to C. gramineum. This primer set could serve as an effective tool in the molecular diagnosis of C. gramineum and has the potential to assist in a better understanding of the host–pathogen interaction.     

E10乙醇汽油在通用小型汽油机中的应用研究

在非道路用车辆中使用的通用小型汽油机未做调整的情况下,试验研究了燃用E10乙醇汽油对发动机动力性、经济性及排放性的影响。试验结果表明,通用小型汽油机燃用乙醇汽油后,动力有所下降,但下降幅度不超过5%;与93#标准汽油相比,中小负荷下乙醇汽油的燃油消耗率有所增加,而大负荷时与汽油基本持平;按照EPA排放法规对其比较排放性,发现乙醇汽油在满足法规要求的情况下,CO有所下降,而HC,NOx有所增长。     

《世界人权宣言》的产生及意义

《世界人权宣言》是联合国制定的第一份系统的纲领性人权文件,由于当时美苏等国不同的人权观以及各国不同的历史文化背景,在制定过程中充满了分歧和斗争。由于各国政府和人民吸取了二战教训,渴望国际人权保障,在人权委员会的努力下《世界人权宣言》最终还是诞生了,并产生了深远的影响。