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。

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.

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)

Clones identification of Sequoia sempervirens (D. Don) Endl. in Chile by using PCR-RAPDs technique

A protocol of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPDs) was established to analyse the gene diversity and genotype identification for clones of Sequoia sempervirens (D. Don) Endl. in Chile. Ten (out of 34) clones from introduction trial located in Voipir-Villarrica, Chile, were studied. The PCR-RAPDs technique and a modified hexadecyltrimethylammonium bromide (CTAB) protocol were used for genomic DNA extraction. The PCR tests were carried out employing 10-mer random primers. The amplification products were detected by electrophoresis in agarose gels. Forty nine polymorphic bands were obtained with the selected primers (BG04, BF07, BF12, BF13, and BF14) and were ordered according to their molecular size. The genetic similarity between samples was calculated by the Jaccard index and a dendrogram was con- structed using a cluster analysis of unweighted pair group method using arithmetic averages (UPGMA). Of the primers tested, 5 (out of 60) RAPD primers were selected for their reproducibility and high polymorphism. A total of 49 polymorphic RAPD bands were detected out of 252 bands. The genetic similarity analysis demonstrates an extensive genetic variability between the tested clones and the dendrogram depicts the genetic relationships among the clones, suggesting a geographic relationship. The results indicate that the RAPD markers permitted the identification of the assayed clones, although they are derived from the same geo- graphic origin.

高温胁迫对虹鳟(Oncorhynchus mykiss)肝脏中抗氧化酶活性和免疫相关基因表达的影响

夏季高温对虹鳟(Oncorhynchusmykiss)网箱养殖会产生极大威胁,高温胁迫会造成虹鳟应激甚至死亡。为探究高温胁迫对虹鳟肝脏抗氧化响应及相关免疫基因的影响,选取虹鳟“水科一号”幼鱼[(24.8±10.0)g]为实验对象,通过连续升温达到高温胁迫条件,筛选出高温敏感组和耐受组,并对它们的抗氧化酶活性、免疫相关基因表达和随机扩增多态性DNA(RAPD)片段化进行比较研究。结果表明:敏感组与耐受组的超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性含量不存在显著性差异(P>0.05),但均显著高于对照组(P<0.05);敏感组的缺氧诱导因子-1α(hif-1α)、白细胞介素-1β(il-1β)以及热休克蛋白10 (hsp10)基因的相对表达量较耐受组的分别高1.8倍、1.28倍和16.5倍(P<0.05);耐受组的热休克蛋白70 (hsp70)基因的表达比敏感个体高1.37倍(P<0.05);RAPD图谱反映出敏感组的DNA损伤比耐受组更严重。高温胁迫对虹鳟幼鱼hif-1α、hsp70和hsp10基因产生强烈的应答性表达上调,且RAPD针对高温耐受个体可有效进行识别。

重庆养殖中华鳖群体遗传多样性的RAPD分析

[目的]利用RAPD标记技术对重庆4个养殖地区中华鳖的遗传多样性进行分析。[方法]选取(潼南、永川、江津、长寿)4个地区养殖中华鳖样本,运用RAPD方法对4个地区的中华鳖遗传多样性与其遗传距离进行了分析。[结果]用20个随机扩增引物得到142个扩增片段,多态片段数量为95个,4个地区的多态位点比例范围为50.78%~64.54%,总多态位点比例为66.90%,总基因多样性为0.227,种群内基因多样性为0.135,遗传分化系数为0.307,基因流为1.127。4个养殖中华鳖群体的遗传距离为0.049~0.151。[结论]重庆4个地区养殖中华鳖遗传多样性较高,且大部分遗传变异存在于种群内,个体间亲缘关系较近,遗传变异度较小。

航天诱变黔草1号高羊茅分子多样性研究

试验以黔草1号为材料,采用RAPD分子标记技术对经航天搭载后的黔草1号高羊茅子代进行遗传多样性分析,探讨空间环境对其遗传多样性的影响。结果表明,利用RAPD引物进行PCR扩增,产生的扩增条带中,多态性条带占83.85%;34份供试材料的相似系数在0.5364~0.7411之间,说明航天搭载黔草1号高羊茅可以造成其遗传物质的变化,诱变后的各株系发生了不同程度的遗传变异,可以从航天诱变后的株系中选择符合育种目标的材料。RAPD聚类分析将供试材料分为七大类。总体来看,具有相似性状的株系基本上能聚在一类,呈现出一定的性状差异。

基于RAPD分子标记的观赏春石斛遗传多样性分析

【目的】利用RAPD分子标记对观赏春石斛的遗传多样性进行分析。【方法】以10份观赏春石斛种质资源为供试材料,采用100条RAPD随机引物进行筛选。【结果】从筛选出来的15条具有良好多态性的引物中扩增得到125个DNA条带,其中80个条带表现出多态性,多态性位点比率为63.89%。聚类分析结果显示,在相似系数为0.37时10份样本分为两大类,在相似系数0.41时可进一步分为两个亚类群。【结论】利用RAPD分子标记可以鉴别观赏春石斛种质资源。

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。

三种兰科植物的保护遗传学研究初探(英文)

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ISSR和RAPD分子标记的15株糙皮侧耳遗传多样性分析

平菇(糙皮侧耳)是我国食用菌栽培主要品种之一,国内各地栽培品种较多,平菇资源的遗传多样性较丰富。为了明确平菇菌株间的亲缘关系及遗传距离,借助ISSR和RAPD两种分子标记技术,对收集到的国内主产区的15个平菇菌株进行DNA遗传多样性分析,为后期平菇适应性试验提供理论依据。通过ISSR和RAPD两种分子标记引物筛选试验,获得9对多肽引物,利用9对引物对15份平菇DNA样品进行多样性PCR扩增,并采用NTSYS-PC分析软件对供试菌株的遗传多样性进行聚类分析。结果显示,参试平菇菌株遗传距离在0.26~0.56;9对引物共扩增产生了102条清晰条带,平均每对引物扩增产生11.3条带;多态性条带90条,多态性条带比例为88%,平均每对引物组合扩增产生10条多态性条带;在遗传距离0.36的水平上,供试的15个平菇品种分为4个类群,P101和P121分别单独分为一个类群,P102、P112、P120、P122、P104、P125、P107、P113、P124归为一个类群,P117、P123、P118、P119归为一个类群。供试材料之间遗传差异大,遗传多样性较丰富,研究结果为本地区平菇产业发展及育种提供新的菌株。

Isolation and Progeny Transmission of Non-Temperature-Sensitive MS-H Vaccine Strains of Mycoplasma synoviae from Temperature-Sensitive MS-H-Vaccinated Laying Breeder Hens

This pilot study reports the vertical transmission and reverse thermosensitivity of the MS-H vaccine strain of Mycoplasma synoviae (MS) by RAPD in commercial breeders and their progeny. At two weeks of age, breeders were vaccinated with the ts+ MS-H strain. At 9 weeks of age, an outbreak of infectious synovitis (IS) was detected in the progeny. Tracheal swab samples were collected from breeders at 24, 39, 48, and 70 weeks of age. At 9 weeks, pullets swab from the elbow joints were collected. RAPD was performed on the isolates at 39.5°C, and the same ts- MS-H strains were identified in the breeder hens and their progeny. Tracheal swabs from breeder hens were negative to MS isolation at 37°C and 39.5°C at 24- and 39-weeks. MS isolation was recovered from tracheal swabs from 9/10 and 10/10 breeders at 48- and 70- week. At 9 weeks of age in the progeny, MS was isolated from tracheal swabs of 10/10 from non-IS pullets. MS was isolated from 9/10 joints samples. The isolates from breeder hens and their progeny showed non-significant differences in five antimycoplasmic MIC100 values;otherwise, enrofloxacin presented a significant difference in MIC100 value (p < 0.05). This investigation demonstrated the reversal of the thermosensitivity, pathogenicity, and vertical transmission of the MS-H strain. Consequently, it is crucial to contemplate the danger of reversing pathogenicity and transmission to progeny when applying the MS-H vaccine strain.

两个广谱抗虫基因源抗性背景DNA的多样性分析

利用８对抗－感大豆叶食性害虫近似等位基因系，１００个随机引物进行ＰＣＲ扩增，在４０６个稳定重现的片段中，有２个在近等基因系间表现多态性；不同抗性来源的两组材料间的多态性ＤＮＡ片段的有／无呈相反趋势，相同抗性来源的材料间的多态性ＤＮＡ片段的有／无也存在差异。由此可以认为，抗性供体ＰＩ１７１４５１，ＰＩ２２９３５８的抗性背景ＤＮＡ的多样性比较丰富，遗传背景相差较大，说明它们的抗性基因来源不同。在进行广谱抗虫育种实践中，为丰富育成品种，尤其是回交育成品种在抗性基因和抗性基因背景ＤＮＡ上遗传多样性，避免其遗传脆弱性，以维持持久的抗性。

青藏高原东缘中国沙棘的克隆多样性及克隆结构

应用随机扩增多态性DNA标记(RAPD markers)对青藏高原东缘中国沙棘的克隆结构、克隆多样性进行了初步的探讨。结果表明,1)14条RAPD引物在8个中国沙棘居群共184个样本中平均形成55.75种条带,平均有15.13种条带在居群内表现出多态性,平均多态条带百分率为27.64%;鉴别出26种基因型,平均每基株形成3.4个分株;2)与应用DNA分子标记研究的其他克隆繁殖能力较强的植物相比,中国沙棘的克隆多样性水平偏低,Simpson’s多样性指数(D)平均值为0.85;Fager’s均匀度指数(E)平均值为0.87;3)克隆结构分析表明,中国沙棘每克隆内分株间的平均距离为1.75m,克隆繁殖的生长型主要为游击型,并且在居群内存在以某一优势基株为主的克隆分布方式;4)居群的定居时间、生境及群落结构等对中国沙棘的克隆多样性有一定影响。