A Molecular Approach to Identification of the Chinese Drug“pu Gong Ying”(Herba Taraxaci)and Six Adulterants byDNA Fingerprinting Using Random Primed PolymeraseChain Resaction(PCR)

DNA fingerprinting among members of the Chinese drug Pu Gong Ying(Taraxacum mongolicum Hand,-Mazz.)and six adulterants of Tu Gong Ying were demonstrated with random-primed polymerase chain reaction(PCR)including arbitrarily primed polymerase chain reaction(AP-PCR)and random amplified polymorphic DNA(RAPD).Distinctive,reproducible genomic fingerprints from DNA from 7 species belonged to Compositae were generated with two long(20 and 24 mer)and one short(10 mer)randomly chosen primers.The Pu Gong Ying can be differentiated from six species of Tu Gong Ying according to the banding pattems of their amplified DNA on agarose gels.The results showed that AP-PCR and RAPD methods can be used for identifying Chinese drugs.Moreover,the Similarity Indexes of the genomic DNA fingerprints showed that Pu Gong Ying and its adulterants are unrelated.Therefore,AP-PCR and RAPD methods can be used for identifying Chinese drugs.

Identification of Porphyra lines using computerized DNA fingerprinting

RAPD (Randomly Amplified Polymorphic DNA) analysis was performed with filaments of 15 Porphyra lines representing four important groups (P. yezoensis, P. haitanensis, P. katadai var. Hemiphylla and P. digospermatangia). Eight stable and repeatable RAPD bands amplified with two primers, OPN-02 and OPJ-18, were selected for the construction of DNA fingerprinting. The RAPD results were scored based on the presence or absence of each of the 8 bands and then converted to computer language expressed with two digitals, 1 and 0, which represented the presence (numbered as 1) or absence (numbered as 0) of each band, respectively. Based on these results, a model DNA fingerprint and a computerized DNA fingerprint were constructed. In the constructed DNA fingerprint, each Porphyra line has its unique fingerprinting pattern and can be easily distinguished from each other. Later, a software, named as PhGI, was designed based on this DNA fingerprinting. It can be used in practical Porphyra line identification.

Identification of Four Gastrodia elata Bl. Variants with DNA Fingerprints

[Objective] This study aimed to investigate the four Gastrodia elata B1. variants at the DNA level. [Method] Primers with good reproducibility, abundant polymorphism and high resolution were selected from the 35 random primers with RAPD marker technology and the amplification results were analyzed. [ Result] By using RAPD technology, DNA fingerprints of four Gastrodia elata B1. variants with abundant polymorphism, good reproducibility and high resolution was successfully constructed; to be specific, each Gastrodia elata B1. variant has its unique fingerprint with primers S29 and S38 ; DNA bands of the Gastrodia elata BI. variants vary significantly, which is contributive to distinguish the four Gastrodia clara B1. variants. [Condusion] This study provided an effective means for the identification and protection of Gastrodia elata germplasms with intraspecific variations.

Application of DNA Fingerprinting Technique in Adulterated Food Identification and Traceability

To further improvc the application of DNA fingerprinting technique in adulterated food identification and traceability, the paper briefly introduced the ap- plication of common DNA fingerprinting techniques, such as species-specific PCR, RAPD, AFLP, ISSR, SSR and SNP in adulterated food identification and traceability.

DNA Fingerprinting of Acer truncatum Clones Using SRAP Markers

[ Objective] This study aimed to establish DNA fingerprints of 23 Acer truncatum clones, thus providing the theoretical basis for selection, classification and identification of A. truncatum varieties. [Method] Sixteen pairs of SRAP primers with rich polymorphism and high specificity were used to establish DNA fin-gerprints. [Result] A total of 223 bands were amplified with 16 primer pairs, including 197 polymorphic bands. Averagely 13.9 loci and 12.3 polymorphic loci were amplified with each primer pair. The average percentage of polymorphic loci reached 88. 34% . [ Conclusion] The classification result drawn by cluster analy-sis is consistent with that obtained based on main characteristics and genetic relationships of A. truncatum, clones. By using DNA fingerprints established with prim-er pairs ME1-EM4 and ME2-EM1, 23 A. truncatum clones can be effectively distinguished, and the confidence probability is greater than 99.99%.

DNA Fingerprinting of Essential Commercialized Medicinal Plants from Pakistan

Development of fingerprints based on DNA markers is necessary for proper identification and standardization of plant species. These techniques are widely used to develop an unquestionable method of plant identification to protect the patents and quality control for industry. In this study, fifteen commercially important medicinal plants of Pakistan were collected from botanical garden of Qarshi Industries (Pvt.) Ltd, Pakistan. The objective was to optimize the extraction of genomic DNA for use in a PCR-based random amplified polymorphic DNA marker approach. The initial protocol used 60 decamers to amplify scorable amplicons;only nine markers produced significant bands in genomic DNA of medicinal plants. These markers generated 51 bands ranging between 250 and 1600 bp. The most important property of genomic markers is polymorphism to enable specific identification;all the used markers showed 100% polymorphism across 15 different plants. Further, six decamers amplified specific bands to reliably identify 8 species. The amplified bands were arranged in a binary matrix and analyzed by DNAMAN version 5.2.2 statistical software. A homology tree was constructed using binary data for nine markers, and four major clusters/clades were observed. The Rose, Mentha and Stevia accessions had shown clear clustering and grouped in major clusters/clads I, II and III respectively. Sixty decamers amplified 51 polymorphic loci in the genomes of 15 commercially valuable accessions. Moreover clear phylogenetic construction was observed in the generation of homolog tree. This protocol could therefore be useful to provide a baseline to authenticate, identify and perform phylogenetic analysis of important medicinal plants used in the Pakistani herbal medicine industry.

50份无花果种质资源遗传多样性分析及DNA指纹图谱构建

为探究国内外无花果遗传多样性并对种质资源进行鉴定,利用15对SSR分子标记引物对50份无花果种质资源从DNA水平上进行遗传多样性分析,并构建DNA指纹图谱。结果表明,15对SSR引物在50份无花果种质资源中共扩增出67个等位基因,等位基因数范围为2~8个,观测杂合度(Ho)和期望杂合度(He)平均值分别为0.5947和0.5703,Shannon’s多态性信息指数(I)平均值为1.0452,多态性信息含量指数(PIC)变化范围为0.2411~0.7353,平均为0.5101,表明无花果种质资源具有丰富的遗传多样性。50份无花果种质资源之间的遗传距离为0~0.68,平均为0.39,与其他种质资源遗传距离最大的为哈代。基于SSR分子标记结果,利用UPGMA方法对50份无花果种质资源进行聚类分析,根据遗传距离可将50份无花果种质资源分为四大类,第Ⅰ类为哈代,第Ⅱ类为斯特拉,第Ⅲ类为梦幻甜蜜,其余无花果种质资源划归为第Ⅳ类。构建了26份无花果种质资源的SSR指纹图谱,可用于品种鉴定,以期为无花果分子辅助育种提供科学依据。

52份枇杷种质资源遗传多样性分析与DNA指纹图谱构建

【目的】丰富枇杷种质资源鉴定方法,为枇杷种质资源发掘和利用提供理论依据。【方法】利用277对EST⁃SSR标记和1对果肉颜色相关InDel标记,对52份国内外枇杷种质资源和国家西南特色园艺作物种质资源圃(成都)枇杷育种与栽培创新团队创制的杂交后代进行遗传多样性分析并构建DNA指纹图谱。【结果】筛选出扩增带型清晰、稳定性和重复性好的9对引物,每对引物扩增条带为2~5条,平均85条;多态性条带比率为40.00%~100.00%,平均71.55%;多态性信息含量(PIC)为0.250~0.999,平均0.685。遗传多样性分析显示,52份枇杷种质资源和创制的杂交后代的平均相似系数为0.791;果肉颜色一致或地理分布相近的枇杷种质资源被聚类到相近的分类单元,并且可以准确地将西班牙、日本枇杷种质资源与其他种质资源进行区分。此外,本研究还构建了52份枇杷种质资源的DNA指纹图谱和色块矩阵。【结论】丰富了枇杷种质资源鉴定的分子标记,为今后发掘分子标记紧密连锁的功能基因或片段提供了理论依据。

基于核心KASP标记的江苏粳稻品种DNA指纹图谱构建

为了建立江苏粳稻品种DNA指纹图谱数据库,本研究利用水稻10K液相芯片对314份来源广泛的水稻种质资源进行SNP基因分型,筛选出一系列高多态性的SNP位点并开发出122个KASP标记。利用122个KASP标记对38份江苏粳稻品种进行检测,以多态性信息量(PIC)>0.3,最小等位基因频率(MAF)>0.2,检出率>0.9为筛选标准,筛选出56个具有高效鉴别效率的核心KASP标记。遗传距离相关性分析结果表明,基于56个核心KASP标记的江苏粳稻品种的遗传距离与基于122个高多态性KASP标记的的江苏粳稻品种的遗传距离相关系数为89.1%,呈极显著正相关(P<0.01),表明56个核心标记可以有效代替122个高多态性标记进行品种鉴定。进一步利用56个核心KASP标记对102份江苏粳稻品种的遗传多样性进行分析,并构建了102份品种的DNA指纹图谱和分子身份证二维码,本研究结果为江苏地区粳稻品种鉴定、选育和种质资源高效利用提供了参考。

基于SCoT分子标记的61份加工型黄瓜种质资源遗传多样性分析及DNA指纹图谱构建

为探明加工型黄瓜种质资源的亲缘关系及遗传多样性,为新品种选育提供理论依据,采用目标起始密码子多态性标记(SCoT)技术,以61份加工型黄瓜种质资源为试验材料,进行遗传多样性分析并构建DNA指纹图谱。结果表明,从100条SCoT引物中筛选出8条扩增条带清晰、多态性高且重复性好的引物;利用筛选出的引物对61份黄瓜种质基因组DNA进行PCR扩增,共扩增出238个位点信息,其中多态性位点227个,平均多态性位点百分比为95.38%;采用NTSYS-pc 2.1软件计算得出供试的61份材料间的遗传相似系数分布在0.588~0.920之间,遗传相似系数最小的材料分别是34号与55号,二者在瓜色、叶片形状及果实形状方面存在较大差异,说明两者亲缘关系最远。基于遗传相似系数对其进行UPGMA聚类分析和树状图绘制,在相似系数0.735处,可将其划分为7个类群,说明61份黄瓜种质材料背景存在差异,但多数地域来源相近、生境相似地区的种质被聚在同一类群。利用2对特异性较大的引物SCoT12和SCoT83构建的DNA指纹图谱可单独鉴别出61份黄瓜种质资源,该图谱可为黄瓜分类与鉴定提供科学依据。

四川省玉米品种DNA指纹检测及遗传多样性分析

为探究四川省玉米品种的遗传多样性,用40对SSR引物检测2021年四川省玉米联合体试验中234个参试品种的DNA指纹,构建参试品种SSR-DNA指纹库,并基于DNA指纹进行遗传多样性分析和DNA指纹分析。结果表明,234个参试玉米品种多态信息整体高于早年吉林省和全国审定玉米品种;鲜食玉米组与其他玉米组遗传距离最远;234个参试玉米品种互为不同品种。2021年四川省参试玉米品种与早年吉林省和全国审定玉米品种相比遗传多样性更丰富,DNA指纹检测通过比例较2014-2019年国家玉米区域试验参试组合有所提高。

50种石斛种质资源的遗传多样性及DNA指纹图谱构建

【目的】分析国内外引种石斛种质资源遗传多样性,揭示不同种质间的亲缘关系,为促进石斛种质的有效利用及保护提供参考。【方法】利用ISSR分子标记技术对杯鞘石斛、金钗石斛、檀香石斛等50种石斛进行遗传多样性和亲缘关系研究分析,并构建DNA指纹图谱。【结果】9条多态性高、重复性好的引物共扩增出112个等位基因,其中,多态性位点102个,多态性比率为93.09%。检测的位点数为9~14,平均为12.44;50种石斛属植物的遗传相似系数为0.15~0.95。聚类分析显示,在遗传相关系数0.70处,可将石斛材料分为12个大组。PCR扩增显示,引物UBC808可将50种石斛完全区分开,基于其扩增的14个清晰的多态性条带构建50种石斛的DNA指纹图谱。【结论】50种石斛种间具有丰富的遗传多样性。

基于SRAP标记的三十三份白及种质资源聚类分析及DNA指纹图谱的构建

以白及(BletillastriataRchb.f.)为试材,基于UPGMA法通过13对SRAP引物组合对6个野生居群的33份白及种质资源进行聚类分析并构建DNA指纹图谱,以期为白及品种鉴定和资源收集提供参考依据。结果表明:13对SRAP引物组合共扩增出182条条带(其中有152条为多态性条带),多态性比率为83.52%,平均Shannon’s信息指数(I)和Nei’s基因多样性指数(H)分别为0.4846和0.3243。6个野生居群根据UPGMA聚类,可分为2个支系。我国华南地区广东茂名、广西百色和华中地区湖南张家界3个白及居群聚为一支;西南地区贵州安龙、云南蒙自和四川峨眉山另外3个白及居群聚为另一支。Manteltest检测表明,物种遗传距离和地理距离间存在显著正相关性(r=0.6246;P<0.001)。4对SRAP核心引物组合构建了33份白及品种的DNA指纹图谱。

Plastid Inheritance in Sweet Potato as Revealed by DNA Restriction Fingerprinting

The inheritance of chloroplast DNA (cpDNA) in sweet potato (Ipomoea batatas Lain.) was analyzed using DNA restriction fingerprinting. The cpDNA fingerprints of hybrids from reciprocal crosses between Xushu18 and AB78-1 were found to be identical to those of their female parents, which reveals that cpDNA of sweet potato is maternally inherited in this intervarietal crossing. This maternal cpDNA transmission pattern does not accord with the putative one based on former cytological studies. The plastid inheritance in Convolvulaceae has been briefly reviewed in this study, and the utility of DNA restriction fingerprinting analysis in the study of plastid inheritance is also discussed.

基于SSR分子标记构建19个杧果品种(系)的DNA指纹图谱

杧果(Mangifera indica Linn.)是一种热带亚热带常绿乔木果树,果实香甜、营养丰富、肉质细滑多汁,富有“热带果王”之美誉。为了加速杧果品种(系)的分子鉴定,本研究根据19份试验材料的表型特征,选择其差异较大的5份杧果材料(红象牙杧、金煌杧、红杧6号、红玉杧、南逗迈4号杧)为模板,进行SSR标记引物筛选,PAGE电泳检测筛选得到多态性较好的引物;将得到的引物再采用HEX、FAM两个荧光标记进行5’端修饰,再通过毛细管电泳技术检测,其扩增片段大小通过数字和字母对每对引物的多态性位点进行编码,构建杧果的DNA指纹图谱和DNA分子身份证。结果表明:经PCR扩增以及PAGE电泳检测,从已开发得到115对SSR标记引物中筛选得到9对稳定性较好、多态性好的引物,引物名称分别为M9、M15、M35、M40、M53、M59、M101、M102、M103。经毛细管电泳检测,结合“数字+字母”处理获得19个杧果品种(系)的指纹图谱代码,如金煌杧的指纹图谱代码为9EE9F8933,黔山杧2号的指纹图谱代码为H6F7564IC,黔山杧4号的指纹图谱代码为3DGE47279。以19份材料的指纹图谱代码和基本信息为载体,通过在线条形码生成程序、二维码生成程序生成其条形码DNA分子身份证、二维码DNA分子身份证。该研究结果为标准化构建杧果DNA分子身份证库奠定基础,为杧果遗传育种、优异基因挖掘提供数据参考。

基于SSR标记的100份国内外小麦种质遗传多样性分析及DNA指纹图谱构建

为了分析国内外引进小麦种质资源遗传多样性,揭示不同种质间的亲缘关系,促进小麦种质的有效利用及保护,本研究以引进的100份国内外小麦种质为材料,利用SSR分子标记进行分子检测,分析供试材料的遗传多样性并构建其DNA指纹图谱。结果表明,利用筛选出的23对SSR引物对100份小麦种质扩增,共获得235个基因位点,其中多态性位点232个,占比达到98.72%。SSR引物检测位点介于6~18个之间,平均等位位点数为10.09个,SSR引物有效等位基因数、Nei’s基因多样性指数和Shannon’s信息指数分别平均为1.35、0.22和0.36。不同小麦种质间的遗传相似系数变幅在0.56~0.96之间,平均为0.73。基于Nei’s遗传相似系数,100份小麦种质可被划分为6类,其中第Ⅵ类群包含的小麦种质数目最多,占到种质总数的62%。同时利用多态性比较丰富的5对SSR引物构建了供试小麦种质DNA指纹图谱。本研究结果可为小麦种质资源的科学利用及有效保护提供一定依据。

安徽地区主栽粳(糯)稻品种遗传多样性分析及DNA指纹图谱构建

以安徽省近年来主栽的21份粳(糯)稻品种为材料,采用国家标准NY/T 1433—2014中推荐的48对SSR引物进行多态性分析,研究品种间的遗传多样性。48对引物中21个SSR标记在21份材料中存在多态性,共检测到59个等位基因,平均每个标记检测到2.81个。遗传多样性分析显示:Shannon’s多样性指数变幅为0.199~1.163,平均值为0.657;Nei’s指数变幅为0.095~0.620,平均值为0.390。多态信息含量(PIC)处于0.091~0.549,平均值为0.331。21个品种间存在一定遗传差异,但等位基因观测数、Shannon’s指数、Nei’s指数均较低,表明21个品种间遗传背景较为狭窄。UPGMA聚类分析表明:21个粳(糯)稻品种的遗传相似系数为0.426~0.954,遗传相似系数在0.62处可将21个品种分为5个类群。根据标记的PIC值和MI值进一步简化标记的使用量,利用7个核心SSR标记构建了21份粳(糯)稻材料的标准指纹图谱,为品种真实性判别、纯度鉴定以及核心种质提纯奠定基础。

DNA fingerprinting of reference strains of Puccinia striiformtis f. sp. tritici in China

The rapid development of molecular biology resulted in the wide application of popula-tion genetics theory to plant pathology.It is the development of efficient molecular DNAmarker systems that gives a great insight into the population genetics of a number offungal plant pathogens such as rice blast fungus Magnaporthe grisea,late blight fungusPhytophthora infestans,barley powdery mildew fungus Erysiphe graminis f.sp.hordeiand wheat blotch fungus Septoria tritici.The knowledge on population biology of plantpathogens is essential for screening and identification of disease resistance genes,for devel-

DNA Fingerprints of Animals Generated With Total Genomic DNA Probes

DNA fingerprinting, developed in 1985, has now become a powerful tool forindividual identification and paternity testing in forensic casework and medicine. It hasalso found wide application to population genetics, evolution, systematics and geneticlinkage analysis in animals, plants and even microorganisms. Conventional DNAfingerprinting requires specific multilocus minisatellite probes-or microsatellite

萱草花药材DNA条形码鉴别与HPLC指纹图谱研究

目的:利用DNA条形码技术对萱草花药材进行物种鉴定,建立其指纹图谱,为萱草花质量控制和鉴定提供依据。方法:提取14批样品的DNA进行序列分析和物种鉴定,并建立高效液相色谱法指纹图谱,采用Waters HSS T3型色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.2%乙酸为流动相,梯度洗脱,流速为1 mL·min^(–1),检测波长为360 nm,柱温为35℃。结果:经DNA条形码鉴定,14批萱草花基原均为黄花菜Hemerocallis citrina Baroni。结合指纹图谱共标定了14个共有峰。结论:DNA条形码和指纹图谱2种方法可对萱草花进行物种鉴定,并能全面反映萱草花药材中的成分信息,可用于萱草花药材的真伪鉴定和批间一致性评价。