适于贵州玉米种质DNA指纹库构建的SSR核心引物筛选

为了筛选出适用于贵州玉米种质DNA指纹库构建的引物,以21份贵州普通玉米自交系和36份糯玉米品种为材料,综合比较PIC值大小、扩增带型清晰度及引物的重复性高低,对87对SSR引物进行筛选,以确定贵州玉米种质DNA指纹库构建的核心SSR引物。结果表明:筛选出的23对和15对SSR引物可分别作为构建贵州普通玉米和贵州糯玉种质DNA指纹库的核心引物。     

浙茄类型茄子品种DNA指纹图谱构建

为了高效快速地区分浙茄类型茄子品种,并为鉴定浙茄品种的特异性和真实性提供依据,利用46份茄子材料,对48个单核苷酸多态性(SNP)标记进行核心SNP标记筛选,并利用筛选到的核心SNP标记,构建了6个浙茄类型茄子品种的SNP指纹图谱。结果表明:48个SNP标记均可转化为KASP标记,对46份茄子材料进行SNP标记的KASP分型,根据分型结果和多态性信息含量,34个SNP标记可作为核心标记,利用这些核心标记构建了6个浙茄类型茄子品种的指纹图谱;通过指纹图谱可以快速高效区分6个浙茄类型茄子品种,为茄子品种鉴定和知识产权的保护提供了理论依据。     

八倍体草莓DNA指纹图谱的构建与初步遗传分析

为对草莓品种进行分子鉴定,从59对SSR引物中筛选出18对多态性好的引物,采用毛细管电泳检测方法对84份八倍体草莓品种进行标记分析,检测每个标记等位变异大小,并进行初步遗传分析。将每个引物对84份草莓扩增的带型按照一定原则进行数字标注,为了简化编码,引物P1~P18分别编号为字母A~R,将引物编号与相应的带型编码进行组合得到该样品在特定引物的扩增产物编码,按照18对引物顺序串联这些编码形成该品种的DNA指纹图谱。结果表明:18对SSR引物共扩增得到多态性条带150个,不同引物扩增到多态性条带数4~17个,共检测到377个带型;各引物的多态性信息含量(PIC)值为0.494~0.908,平均值为 0.738;利用18对SSR引物共构建了84个草莓品种的DNA指纹图谱。聚类分析结果表明:在遗传相似系数为0.99时能将所有品种区分开,一些来源相同的品种被优先聚在一起。综上,本研究成功构建84个草莓品种的DNA指纹图谱,这些品种遗传关系较近。     

DNA amplification fingerprinting and marker screening for pseudo-testcross mapping of flowering dogwood ( Cornus florida L.)

DNA amplification fingerprinting (DAF) with arbitrary oligonucleotide primers was used to study genetic relationships between cultivars of flowering dogwood (Cornus florida L.), evaluate extent of plant hybridization, and generate markers in pseudo-testcross mapping at the intraspecific level. Modified Taguchi optimization methods defined a robust DAF system based on high annealing temperature (48–52 °C) and primer concentration (typically 8 μM) that was used to study genetic diversity of representative dogwood cultivars and hybrids. Phenetic analysis using cluster and numerical methods showed that: (1) cultivars were relatively conserved at the genetic level; (2) their hybridization could be identified in the F1 progeny in the absence of phenotypic or physiological markers; (3) several cultivars grouped according to their recorded ancestry; and (4) dogwood anthracnose-resistant lines originally selected in Catoctin Mountain Park (Maryland) grouped separately from those of southern origin. The DAF protocol was also tested in pseudo-testcross mapping of dogwood at the intraspecific level. A preliminary screening of parents ‘Pink Sachet’ and ‘Fragrant Cloud’ and 7 F1 segregants with 22 octamer primers produced 703 amplified loci, 30 and 39 of which were male and female markers segregating at 1:1 ratios with 98.6% confidence levels in pseudo-testcross configuration. Overall results show that DAF generated markers very efficiently (3 per primer) despite the close relatedness of parental dogwood cultivars. This study constitutes the basis for a future genetic linkage mapping and marker-assisted selection (MAS) effort initially targeted to control important fungal diseases in dogwood. This revised version was published online in July 2006 with corrections to the Cover Date.     

棉花AFLP银染技术及品种指纹图谱应用初报

本文介绍了一种适合于棉花研究的ＡＦＬＰ银染方法,并利用该方法获得了几个棉花品种的指纹图谱     

Low level of polymorphism detected by SSR probes in bread wheat

In-gel hybridization patterns were studied in a set of nine diverse bread wheat ( Triticum aestivum L. em. Thell) genotypes using 23 simple sequence repeat (SSR) probes in combination with 14 different restriction enzymes. Multilocus fingerprints due to SSR probes, shown earlier to be characteristic of a majority of plant genomes, were not obtained and only a very low level of polymorphism was detected when using as many as 142 probe-enzyme combinations. The hybridization of a prominent solitary high molecular weight fragment (> 23 kb) with a number of SSR probes suggested the presence of these SSRs (microsatellites) within the long stretches of repeated DNA sequences. This indicates that the genome of bread wheat differs from that of other plants in the organization and distribution of SSRs and that SSR probes detect very little polymorphism.     

基于SSR标记的白化和黄化茶树品种遗传多样性分析及指纹图谱构建

利用62对SSR引物对16个白化、黄化茶树品种资源的遗传多样性进行了分析,初步明确了不同白化、黄化品种的遗传结构,以及SSR标记在白化、黄化品种鉴定上的适用性,为此类茶树品种资源的鉴定评价提供了理论依据。经过对引物筛选和扩增条带的分析,结果显示：具有多态性的55对引物中,共检测出169个等位基因,每对引物检测出的等位基因数为2~5个,平均为3.07个;多态信息含量（PIC）和Shannon信息指数（I）的平均值分别是0.40和0.79;169个等位基因出现频率在3.12%~96.88%之间;16个参试品种的遗传距离在0.086~0.532,品种间遗传结构差异明显;当D≈0.18时,可将16个品种划分为3类,其中大部分亲缘关系相近或地理位置一致的品种聚为一类。此外,笔者根据不同引物的等位基因带型构建了16个白化和黄化茶树品种的分子指纹图谱,并筛选出3对核心引物（TM156、TM508、MSG0380）用于不同白化、黄化茶树品种的鉴定。     

茶树DNA分子指纹图谱研究进展

分子指纹图谱直接反映生物个体在DNA水平上的差异,可有效鉴别茶树品种,保护品种权。本文概述了指纹图谱的分类、概念和特点,近年来国内外7种分子指纹图谱方法在茶树中的研究进展,以及指纹图谱在茶树种质资源与育种研究中的应用,探讨了我国茶树分子指纹图谱研究中存在的一些问题,并由此提出构建茶树标准指纹图谱的设想。     

新陆早棉花品种DNA指纹图谱的构建及遗传多样性分析

以新疆2013年前审定的51个新陆早常规棉花品种为材料, 从5000对SSR引物中筛选出多态性高、稳定性好、且定位在棉花26条染色体上(每条染色体上选择2~3对)的75对核心引物,检测到多态性位点226个, 每个标记检测到的基因型位点数在2~12之间, 平均为3.01个;引物多态信息量(PIC)值介于0.0799~0.8752之间, 平均值为0.6624。结果显示, 在51份新陆早棉花常规品种中, 可利用特征引物将21份品种一次性区分开。利用40对引物可以完全区分开新陆早51份常规品种, 并构建供试品种的指纹图谱。同时利用NTSYS-pcV2.10软件聚类分析表明, 51个新陆早棉花品种遗传相似系数变化范围为0.4269~0.9873, 平均值为0.7071, 说明新陆早棉花品种之间遗传多样性较狭窄;遗传相似系数矩阵和聚类分析将51个新陆早品种分为4大类型, 与原品种选育系谱高度吻合。