RAPDs和RFLPs分析甘蓝型杂交油菜亲本的遗传多样性

利用RAPD和RFLP分子标记技术对甘蓝型杂交油菜亲本遗传多样性的分析结果表明 :(1) 2 0个亲本间具有丰富的遗传多样性 ,4 0个引物扩增出 2 77条多态性带 ,其中 2 1条带为 10个亲本所特有 ,12个探针得到了 117条多态性杂交带 ,其中 7条带为 5个亲本所特有 ;(2 )不育系与恢复系之间的遗传差异大于不育系内和恢复系内的遗传差异 ;(3)依RAPDs与RFLPs估计的亲本遗传多样性存在有一定的差异 ,这种差异由估计时所用多态性带的数目不同而引起 ;(4)NJT聚类方法将 2 0份亲本分为 3组 :不育系组、恢复系组和混合组。杂交种“秦油 2号”和“华杂 3号”的双亲都被聚在不同的组内。

旱麦草属种质资源的随机扩增多态性DNA(RAPDs)分析

利用３１个１０ｂｐ随机引物对来自我国新疆和中东地区的 １１份早麦草属种质资源材料以及两个普通小麦亲本和１个普通小麦与东方旱麦草的远缘杂种后代进行了ＭＰＤ分析。对扩增形成的３２１条谱带进行的研究发现，该属植物在新疆地区具有较丰富的遗传多样性，同时还将１个根据形态性状定名为光穗旱麦草的样品修正为东方旱麦草。研究还揭示出早麦草属与普通小麦之间有明显的遗传分化，并发现四倍体光穗旱麦草的两个基因组分别来自二倍体光德旱麦草和毛穗旱麦草。结合前人研究结果，建议将光穗旱麦草的两种细胞类型分列为两个独立的物种，将３个二倍体物种──光穗旱麦草、毛穗旱麦草和旱麦草的基因组符号分别定为Ｆ、Ｆ和Ｆ同时还提出了三者的起源演化关系。

Analysis of Genetic Structure of Natural Populations of Castanopsis fargesii by RAPDs

Genetic diversity and population genetic structure in 188 individuals from five natural populations of Castampsis fargesii Franch. were studied by RAPD markers. Three hundred and eighty-five loci were identified with 41 oligonucleotide primers, out of which 157 loci were polymorphic and accounted for 40.78% of total genetic diversity at species level. Shannon's indices of diversity (I) and Nei's gene diversity ( h) were 0.459 7 and 0.296 at the species level, respectively. The result showed that genetic variation of C. fargesii populations mainly existed within populations. Genetic differentiation (Hsp-hpop)/Hsp estimated with Shannon's index of diversity and coefficient of gene differentiation (Gst) were 0.047 6 and 0.042 9 respectively, which were confirmed by the analysis of molecular variance (AMOVA). Therefore, it is apparent that within-population variation accounted for 94.97% and among-populations variation accounted for only 5.03% of the total genetic diversity. AMOVA also indicated that there was significant differentiation among populations as well as among individuals within a population.

Isolation and Chromosomal Mapping of a Corn B Chromosome Specific RAPDs

B染色体存在于多种动植物中 ,具有很多独特的性状。B染色体与正常染色体在DNA组成方面十分相似 ,寻找B染色体特异序列一直是B染色体研究的难点和热点。通过对含有和不含有B染色体的两种玉米 (ZeamaysL .)基因组进行了RAPD分析 ,筛选到一个B染色体特异性分子标记B480。该标记与玉米的自主复制起始序列ARS1和ARS2同源 ,特别是该序列中的 2 5bp出现在多种模式生物基因组中。FISH的结果显示 。

用RAPDS方法鉴别桉树无性系

前言:Orange Free州大学林木病理合作项目的Brenda Wingfield博士报道了一种正在研究中的新的确认桉树特征的指纹方法。她认为如果要有效地进行林木无性繁殖,需要采取先进的方法以确定无性系的特征。这项研究很快被研究多种农作物的专家们所认同。

Somaclonal variations of Soybeans (<i>Glycine Max.</i>L. Merr) stimulated by drought stress based on random amplified polymorphic DNAs (RAPDs)

In soybeans, drought stress causes 50% yield losses. Breeding for drought tolerance in soybeans has been widely developed using various methods, among which is polyethylene glycol (PEG-6000) induction to simulate drought in vitro. In a previous experiment, three somaclones with different levels of tolerance were generated. The objectives of this research were to determine the RAPD patterns of those somaclones and to investigate the correlation of the RAPD patterns to the drought tolerance characteristics. The results showed eleven RAPD primers capable of amplifying the DNA genome of soybeans, among which four primers were monomorphic and seven were polymorphic. Two of the polymorphic primers, OPK7 and OPK12, are capable of differentiating medium tolerance traits from other traits. Bands that are specific for medium tolerance against drought were 450 bp and 650 bp in size, generated by the OPK7 primer, and the band of 2000 bp, generated by the OPK12 primer. However, there was no band capable of differentiating between sensitive and tolerance varieties/lines, although some changing of the DNA sequence was detected in this research. This indicates that there are other factors responsible for the expression of drought tolerance.

Assessment of Genetic Variation and Distribution Pattern of Thalictrum petaloideum Detected by RAPDs

Random amplified polymerphic DNA(RAPD)method was applied to assessg enetic variation and population structure of Thahctrum petalotdeum L(Ranunoulaceae)，Two hundred and forty-six individuals from 11 populations of the species were investigated by RAPD profiles Twenty selected RAPD primers generated 125 bands．in which 120 were polymorphic Ther esults revealed a high level of genetic variation(ercentage of polymorphIc bands(PPB was 96％．Nei’s gene diversity(りwas 03502 and shannon’s information index(I) was 0．5199 at the species level) The differentiation among the populations was high(Gst=0．3511)in this species．Result of analyzing of molecularvariance(AMOVA)showedthat38．88％of genetic variance was found among the populations Positive correlation withr r=01945(P=00002)was found between genetic distance and geographic distance amongpo pulations Two populations distributed in the drainage basin of YanELz River affined genedcally and formed one clada and the rest nine populations formed the other clade in both unweighted pair-group method using arithmetic average(UPGMA)trees made by two different method different methods. It was yen/clear that these two populations were very special, andmust be closely related in history, despite the fact that they now share quite weak link to the restpopulations through gene communication.

Comparison of Genetic Diversity of the Germplasm Resources of Confectionary Sunflower (Helianthus annuus) in China Based on RAPDs and AFLPs

RAPDs and AFLPs were used to determine the genetic relationships among 23 elite cultivars of confectionary sunflowers (Helianthas annuus) from different districts in China. Both approaches uniquely fingerprint each of the accessions. Twenty-six RAPD primers resulted in a total of 192 strong DNA fragments, ranging from 0.26 kb to 1.98 kb, among which 165 (86.12%) were polymorphic. The average number of DNA band produced by each primer was 7.38. A total of 576 AFLP markers were produced with 8 primer combinations, ranging from 100 bp to 500 bp, and 341 polymorphic bands (59.20%) were revealed. The polymorphism rate was 76.00% and the average bands amplified by per primer combination were 72. Effective number of alleles per locus of RAPD marker (1.76) was larger than that of the AFLP marker (1.65). The mean PIC value of AFLP markers (0.38) was lower than that of the RAPD markers (0.41), but AFLP marker had much higher Ai value (38.52) than RAPD marker (6.38). Genetic similarities from RAPD data ranged from 47.84% to 82.06% and the average Nei's coefficient was 0.649 5; the Nei's coefficient of similarity from AFLP data ranged from 54.15% to 83.52%, and the average Nei's coefficient was 0.688 4. However, standard deviation (SD) of RAPDs was 0.13 but the SD of AFLPs was 0.08. In general, the RAPD data gave lower similarity values and higher SD values than those based on the AFLP analysis. The correlation coefficient between the two genetic similarity matrices was 0.51, revealing the estimations of genetic relationship provided by the two marker systems were only moderate. However, cluster analyses of RAPD or AFLP data divided the 23 sunflower genotypes into identical 3 groups.

Genetic diversity of hybrid durian resulted from cross breeding between <i>Durio kutejensis</i>and <i>Durio zibethinus</i>based on random amplified polymorphic DNAs (RAPDs)

One of the ways to improve the quality of Indonesian Durian is by utilizing germplasm diversity. Durio zibethinus is the most cultivated durian in Indonesia, whereas Durio kutejensis is a unique durian cultivar which has golden yellow fruit flesh without smell. Crossbreeding of those two cultivars, in order to generate superior Durian cultivars has been done. Genetic diversity of durian generates from cross breeding between D. kutejensis and D. zibethinus was identified in molecular level using RAPD technique. Among 20 primers used in this study, 5 primers: OPA-02, OPA-03, OPA-08, OPA-10 and OPA-13 were capable of differentiating both the parents and the hybrids. RAPD analysis resulted in genetic diversity of hybrid Durian with family relationship of 0.59%-0.1%. Hybrids UB1, UB5, UB13, UB19, UB21, UB7 and UB35 have similarity value of 0.81% with their parent DRCK, whereas hybrids UB8, UB10, UB18 and UB17 have similarity value of 0.70% with their parent DRCM1. Hybrids UB2, UB16 and UB22 belong to one group with similarity value of 0.67%. Three hybrids lines UB2, UB16 and UB22 show the highest distance to both parent. The rest of the hybrids lines grouped into similar cluster to the parents D. kutejensis, whereas the other parent (D. zibethinus) belong to different cluster separated from all other hybrid lines and parents.

采用形态学性状和RAPDs标记评价印度四倍体和二倍体棉花的遗传多样性

印度是世界上最大的棉花生产国，但生产力仅为300kg／ha，远低于世界的平均水平544kg/ha。过去50年，印度经历了由二倍体棉花向四倍体棉花的转变。二倍体棉花具有的几个有用性状能转育到目前栽种的四倍体棉花中。特定转育材料间有用性状的筛选取决于亲本材料的遗传多样性。在分子标记(RAPD)

基于RAPD和ISSR标记的云芝种质资源遗传多样性分析及SCAR标记建立

为了探究野生云芝(Trametes versicolor)种质资源遗传背景,基于RAPD、ISSR两种分子标记技术对18个不同种源的云芝菌株进行鉴定及遗传多样性分析,并建立稳定性更好的SCAR标记。结果表明RAPD和ISSR引物均能扩增出较多条带,多态率分别为100%和97.4%,说明RAPD和ISSR可以检测到供试菌株丰富的遗传多样性,能够对云芝菌株进行有效的区分。南方的长沙居群和宁乡居群的NPL、PPB、Na、Ne、He、I相近且偏高,北方的山东、辽宁和吉林居群偏低。从聚类结果看,整体上呈由南往北聚类的规律。获得的SCAR标记验证结果显示可能与云芝子实体边缘厚度基因相关。该研究完善了云芝遗传物质的分子标记,可为后续云芝品种选育、性状关联基因调控提供一定的研究基础。

基于RAPD的MCID法快速鉴定不结球白菜品种和遗传多样性分析

不结球白菜是长江中下游地区蔬菜周年供应的主栽品种,在种质资源保护和开发利用过程中,品种鉴定的难度随着市场上品种的增多而增加。利用基于DNA分子标记的人工绘制品种鉴别图(manual cultivar identification diagram,MCID)方法对笔者所在单位收集和保存的34份不结球白菜种质材料进行鉴定,同时进行NTSYS-pc遗传多样性分析。结果表明,利用筛选出的5对RAPD引物扩增出的多态性谱带即可构建34份不结球白菜的品种鉴定图,该图可以快速地筛选出某品种鉴定所需要的引物及需要参考的特征性谱带,具有较高的可操作性和实用性。聚类结果表明,34份不结球白菜种质材料与品种的聚类模式基本符合其农艺性状表现和遗传背景。研究结果可为不结球白菜品种鉴定和种质创新提供了科学依据,对不结球白菜种质资源的保护、研究及品种的早期鉴定具有重要意义。

基于PCR技术的松材线虫分子检测技术研究进展

松材线虫是世界性检疫对象,并对我国林业造成了严重的危害。由于线虫个体较小且鉴定特征单一,利用传统的形态学鉴定较为困难,因此客观的分子生物学鉴定方法显得尤为重要。本研究总结了近些年主要的松材线虫PCR检测技术及其与拟松材线虫的区别,阐述了常规PCR技术、PCR-PFLPs检测技术、RAPD和SCAR分子标记技术以及荧光PCR技术等各类检测技术的特异性序列、片段大小和应用场景,并探讨了各检测技术的优缺点,为后期研究提供参考。

基于RAPD技术研究脉冲强光对大肠杆菌胞内DNA的影响

本实验基于RAPD技术,研究脉冲强光对大肠杆菌DNA的影响,探讨脉冲强光杀菌的机制。方法 使用传统CTAB法,对大肠杆菌和铜绿假单胞菌的DNA进行提取和纯化。通过研究相关文献资料,挑选出适合的特定引物序列,进一步采用RAPD技术对细菌DNA进行扩增处理,并采用琼脂糖凝胶电泳和凝胶成像技术检测DNA结构的细微变化。结果 CTAB法提取大肠杆菌DNA效果良好,纯度高,无蛋白质污染。RAPD结果显示,脉冲强光可以引起大肠杆菌高分子量区域的条带减弱或消失,但是在250~3000 bp分子量区域没有明显的新增条带。结论 脉冲强光会导致大肠杆菌的DNA分子发生断裂或者降解,本实验结果为探讨脉冲强光杀菌的机制提供了基础。

江西省烟草赤星病菌RAPD遗传多样性分析

【目的】为明确江西省烟草赤星病菌Alternaria alternat的遗传多样性,开展了相关研究。【方法】运用RAPD分子标记法对江西省6个主要烟区赤星病分离出的37个菌株进行了遗传多样性、遗传结构与分化及种间遗传变异分析。【结果】其中6个不同地理种群的Nei’s基因多样性指数(He)为0.1618~0.3209,Shannon信息指数(I)为0.2419~0.4785,表明不同地理来源种群之间的链格孢菌具有丰富的遗传多样性;遗传距离分析表明,兴国和信丰的菌株遗传距离最小,亲缘关系较近,石城和黎川的菌株遗传距离最大,亲缘关系较远;居群每代迁移数(Nm)为1.8171,表明不同地理来源的种群间存在一定程度的基因流动;利用NTSYS软件按UPGMA法聚类分析,当相似系数划定为0.56时,可将37个菌株划分为10个类群,37个菌株基本上以地理来源聚类到不同分支,表明菌株间的遗传多样性受其地理来源影响明显。【结论】利用RAPD分子标记法明确了江西省主要烟区烟草赤星病菌Alternaria alternat的遗传多样性,为江西省烟区选育抗赤星病品种及其综合防治提供措施奠定理论基础。

RAPD分子标记与玉米杂种产量优势预测的研究

以24个优良玉米自交系按NCⅡ（11×13）设计组配成143个单杂交种为材料，利用RAPD分子标记技术研究玉米杂种优势群划分，遗传距离（GD）与特殊配合力、杂种产量、杂种产量优势（MH）的关系。结果表明：（1）RAPD技术可用于玉米杂种优势群划分。（2）亲本遗传距离与杂种产量优势、杂种产量、特殊配合力有一定相关关系，但决定系数很小，分别是10％、10％、15％，利用RAPD技术预测杂种优势、杂种产量作用有限，应进一步研究与杂种优势有关的数量性状位点（QTL），从而使育种家预测高产组合成为可能。

转基因抗虫棉品种(系)的遗传多样性初步研究

利用简单重复序列 (SSRs)和随机扩增多态性DNAs(RAPDs) 2种分子标记对来自我国各个棉区的 30份陆地棉转基因种质材料进行了遗传多样性分析 ,材料包含有我国的 3种转基因类型。利用NTSYS -pc 2 .10统计分析软件 ,采用Jaccard’s相似系数UPGMA法进行聚类。结果表明 ,3类转基因材料大致分为不同的 2类 ,而且不同棉区的转基因材料交叉分布。相似系数分析表明 ,我国转基因材料的遗传多样性相对较为丰富。

A Preliminary Study on Conservation Genetics of Three Endangered Orchid Species

采用随机扩增多态DNA(RAPD)分析研究了中国 3种珍稀濒危兰科植物硬叶兜兰 (PaphiopedilummicranthumTangetWang)、麻栗坡兜兰 (P .malipoenseS .C .ChenetTsi)和独花兰 (ChangnieniaamoenaChien)的遗传多样性与群体遗传结构。 12个RAPD引物在 2种兜兰中共扩增出 131条带。对 4个硬叶兜兰群体的检测表明其物种水平的多态条带百分率 (PPB)为 71.6 % ,Nei的基因多样度 (h)为 0 .2 171,Shannon多样性指数 (I)为 0 .330 1;4个群体的平均多样性水平为PPB =45 .2 % ,h =0 .145 7,I =0 .2 2 0 4,低于远交兰花的平均水平。在总遗传变异中 ,群体间遗传变异占 2 0 .31% ,略高于远交物种的平均水平。在物种水平上 ,麻栗坡兜兰的PPB为 49.5 % ,h为 0 .1174,I为0 .176 4,均大大低于硬叶兜兰。对 11个独花兰群体采用 16个RAPD引物共扩增出 119条带。物种水平PPB =76 .5 % ,h =0 .1941,I=0 .30 5 8;在群体水平上 ,上述 3个指标的平均值则分别为 37.2 %、0 .1197和 0 .1810 ,均低于远交兰花的平均水平。群体间的遗传变异占 45 .2 7% ,遗传分化明显高于远交物种的平均水平。导致 3个物种遗传多样性偏低而群体间遗传分化较高的主要原因在于人为的过度采挖和生境的片断化。

基于RAPD技术对大曲芽孢杆菌近似菌株的分类研究

大曲是浓香型白酒酿制过程中最主要的微生物源,大曲中微生物的类型、丰度、新陈代谢活动等均会对浓香型白酒品质产生很大的影响。为了探究大曲中芽孢杆菌的分类及多样性,利用菌株随机扩增多态性DNA标记(random amplified polymorphic DNA,RAPD)分型和16S rDNA序列分析技术,对已分离出的芽孢杆菌进行RAPD分型和16S r DNA序列分析。结果表明,37株芽孢杆菌中有22株枯草芽孢杆菌、9株蜡样芽孢杆菌、3株地衣芽孢杆菌,其中枯草芽孢杆菌又可细分为8类,蜡样芽孢杆菌可细分为3类,地衣芽孢杆菌可分为3类;RAPD分型可以为大曲中芽孢杆菌的快速分型及后续的分类学研究提供理论依据,为研究芽孢杆菌在白酒酿造中的作用奠定基础。