中国李RAPD的优化反应体系及其在品种鉴定中的应用

以5'-GACCGCTTGT-3'为随机引物,以奉化李(Prunussalicinacv.Fenghuali)为试材,对中国李RAPD反应体系进行了优化研究,结果表明:25μL反应体系中,TaqDNA聚合酶、Mg2+、引物、模板DNA和dNTPs5种主要成分的适宜浓度或用量分别是:1.0U、2.5mmol/L、20ng、40ng和0.2mmol/L。利用该优化反应体系,用10个随机引物,构建了5个中国李品种的RAPD指纹图谱;并以共有谱带率对这些品种遗传关系进行了分析,5个中国李品种的共有谱带率平均为33.2%,变异幅度为20.1%～44.9%。     

Genotyping of Mycobacterium bovis by geographic location within Mexico

The spacer oligonucleotide typing (spoligotyping) method was used to differentiate 62 Mycobacterium bovis isolates obtained from tissues with macroscopic lesions typical of tuberculosis in dairy cattle from different regions of Mexico. Our purpose was to see if a strain from one region was genetically different from those of other regions (with the long-term aim of doing molecular trace back of isolates obtained in the laboratory). Results from the genetic analysis indicate that M. bovis isolates cannot be grouped by geographic location due to a wide range of genetic types involved in dairy cattle infections. Isolates even from the same herd showed different spoligotypes but some isolates from different region had similar genetic patterns. Genetic typing without epidemiologic information does not seem to be a plausible method to trace back animals to source of origin to detect and eliminate sources of infection.     

番茄品种SSR标记鉴定技术研究

以16个番茄品种为材料,从200对番茄SSR引物中筛选出20对适合用于番茄品种鉴定的引物,用该20对引物对66个番茄品种进行分析,共检测出52个等位基因变异,等位基因变异范围为2~5个,平均每个位点产生2.6个等位基因变异。通过遗传聚类方法对引物组合的品种鉴别能力进行分析,结果显示仅用9对引物构成的引物组合就可以对66个番茄品种进行有效鉴别,该9对引物组合构建的SSR-DNA指纹图谱可作为品种的遗传标签用于品种真伪鉴定。     

直接扩增甜瓜小卫星DNA指纹图谱

以小卫星DNA YNZ22核心序列为引物,甜瓜DNA为模板,在甜瓜上研究了直接扩增小卫星DNA(DAMD)的优化反应体系,结果表明:在20μL的反应体系中,5种主要成分Taq DNA聚合酶、Mg2 、引物、模板DNA和dNTPs的最适浓度分别为1U,2.5 mmol/L,0.5 mmol/L,30 ng,5 mmol/L。在优化的DAMD-PCR反应体系下,利用该引物在28个甜瓜品种上构建了直接扩增小卫星DNA(DAMD)的指纹图谱,分析结果表明,该引物在28个甜瓜品种上共扩增出13位点,其中9位点具有多态性,多态性条带比率为69%,可一次性从28个甜瓜品种中鉴别出其中的21个,鉴别率高达75%。     

Database derived microsatellite markers (SSRs) for cultivar differentiation in <Emphasis Type="Italic">Brassica oleracea</Emphasis>

Fifty-nine Brassica oleracea cultivars, belonging to five botanical varieties, were evaluated for microsatellite (SSR) polymorphisms using 11 database sequence derived primer pairs. The cultivars represented 12 broccoli (Brassica oleracea var. italica), ten Brussels sprouts (B. o. var. gemmifera), 21 cabbage (B. o. var. capitata, including the groups white and red cabbage), six savoy cabbage (B. o. var. sabauda), and ten cauliflower (B. o. var. botrytis) cultivars from 13 seed suppliers. The 11 primer pairs amplified in total 47 fragments, and differentiated 51 of the cultivars, whereas the remaining eight cultivars were differentiated from the rest in four inseparable pairs. All SSR markers, except one, produced a polymorphic information content (PIC value) of 0.5 or above. The average diversity for all markers within the tested material was 0.64. There was no major difference in the diversity within botanical varieties and groups. The cluster analysis and the resulting dendrogram showed that the cultivars tended to group within these taxonomic units. The present study substantiates the use of microsatellite markers as a powerful tool for cultivar differentiation and identification in vegetable brassicas.     

Multi-parameter fingerprinting of sediment deposition in a small gullied catchment in SE Australia

The determination of relative contributions of potential sediment sources is an important step in the development of management strategies to combat soil erosion. In a 1.2 km² gullied catchment in southeastern New South Wales, multi-parameter fingerprinting of sediment deposited in successive downstream pools has identified gully walls as the dominant sediment source when the grazed pasture surface was the only other potential source. The median fractional contributions remained relatively steady in the successive downstream pools, with the gully walls responsible for between 90% and 98% of the pool sediment. This result was achieved despite the ratio of the source areas varying considerably between successive nested subareas. Reliability bounds on the predictions, accounting for limited sampling of sources, were well constrained and varied between 5.4% and 13.8%. Downstream of an unsealed road crossing, sediment from the road source dominated the pool sediments such that contributions from the pasture surface and gully sources could not be determined.     

Use of Microsatellite Polymorphisms to Develop an Identification Key for Tunisian Apricots

Microsatellite polymorphisms in 54 apricot landrace cultivars were identified by using 26 Prunus microsatellite primers. Samples of apricot cultivars were collected in eight growing regions in Tunisia ranging from the north to the south of the country and from the sub-humid to the saharian areas. The primers revealed 103 alleles and 155 different genotypes among the 54 apricot accessions. The most polymorphic microsatellite loci were used to develop an identification key. Five microsatellite primers and the 28 resulting alleles were sufficient to discriminate among all 54 cultivars. These results are discussed in the context of cultivar nomenclature, geographic origins, and the comprehensive fingerprinting of Tunisian apricot collections.     

郑抗系列无籽西瓜品种SSR指纹图谱的构建

为实现无籽西瓜品种的快速、准确鉴定,用23对西瓜核心SSR引物分析了15份无籽西瓜品种的DNA指纹。23对核心引物中有21对具多态性,共扩增出58个基因型,平均2.71个,平均多态性信息量(PIC)为0.36。10对引物在9个品种上具有特征谱带,组合PIC值0.4的8对引物可以将参试的15个无籽西瓜品种一一区分开,并利用这8对引物构建了参试品种的数字指纹图谱。研究表明,西瓜核心SSR引物适合用于构建无籽西瓜品种的DNA指纹,将为无籽西瓜品种真实性鉴定和知识产权保护提供有效的技术依据。     

指纹技术识别泥沙来源:不确定性研究进展

泥沙指纹技术是近年来研究流域泥沙来源与动态变化的一项重要技术方法,其主要局限性在于诸多的前提假设与实践条件的差异带入了一系列不确定性。泥沙从源到汇的迁移过程受自然环境和人为干扰的影响,其源地特征、迁移路径以及泥沙性质的潜在变化等均是指纹技术应用的不确定性因素,而采样方案设计、指纹选择方法、混合模型应用的一些主观因素进一步增加了不确定性的来源。不确定性分析是泥沙指纹技术的重要组成部分,是评估泥沙来源指纹识别结果可靠性的主要依据。本文综述了泥沙指纹技术的诸多不确定性问题以及主要应对方法的研究进展,总结了当前不确定性定量评估的研究现状并提出了未来的研究展望。     

甜菜SCoT核心引物的筛选

【研究目的】为了筛选出适合甜菜品种遗传多样性分析的SCoT引物,为SCoT引物得以在甜菜中进行深入应用奠定基础,本研究甜菜以8个来自不同国家的甜菜品种为材料对80条SCoT引物进行扩增【方法】扩增方法采用SCoT-PCR最优反应体系包含2.0 μL的10×PCR buffer（含Mg2+）、10 ng的DNA、0.5 U的Taq DNA聚合酶、10 μmol/L的引物2 μL以及0.1 μL的dNTPs (2.5 mmol/L each)。【结果】结果从80条SCoT引物中筛选出20条多态性高的引物,这20条引物最多扩增出16条多态性条带,最少扩增出2条多态性条带,扩增总条带155条,其中多态性条带为134条,多态性条带的百分比为86.5%。【结论】通过对核心引物的有效性验证,表明筛选出的20个核心引物非常适合用于甜菜指纹图谱的构建。