RAPD分子标记及其在作物遗传育种中的应用

ＲＡＰＤ分子标记是一种新型的分了遗传标记,其原理是采用１０个碱基的随机引物,以基因组ＤＮＡ为模板进行ＰＣＲ随机扩增。其优越性在于其方法简单;分析中的花费少,用时短;分析所需的ＤＮＡ用量也不多等。本文着重介绍以电泳技术和ＰＣＲ扩增技术为核心的分子标记以及在农作物遗传育种中的应用。     

偏分离分子标记的作图方法

对取自MAPMAKER软件小鼠F_2群体(含333个体)的5个RFLP连锁标记数据作了共显性分子标记偏分离的分析。先确定选择类型的方程组(配子或合子),随后采用Newton-Raphson迭代法估算标记间的重组值。在构建分子标记遗传图谱时,如果两个相邻标记均存在偏分离,最好采用纳入偏分离因子的估算方法。在估计F_2群体标记间偏分离重组距离上,用连续x~2检测方法比传统x~2检测更为准确。     

Comparative studies on the nuclear polyhedrosis viruses of Lymantria monacha and L. dispar

Immunodiffusion and tube precipitation tests, polyacrylamide gel electrophoresis of virus polypeptides, and cross-transmission experiments suggest that two nuclear polyhedrosis viruses, one from Lymantria monacha and one from L. dispar, are partially related to each other, but not identical. The virus particle proteins seem to be more specific than the polyhedron proteins.     

苏铁属植物RAPD反应体系的优化及部分种类亲缘关系的探索

通过优化苏铁属植物的RAPD反应体系,进而探讨苏铁属部分种类的亲缘关系。结果表明,Mg~(2+)、dNTP、Taq酶及随机引物浓度在RAPD反应中有重要影响,而模板DNA浓度有一个很大的适应范围。适合苏铁属植物RAPD分析的反应体系:20μL反应体系中,含有10 mmol/L Tris-HCl(pH8.3)、50 mmol/L KCl、2.0mmol/L MgCl_2、200μmol/L dNTP、0.3μmol/L随机引物、模板DNA 50ng、Taq酶1.0 U。聚类分析结果基本反应了苏铁属各个种间的亲缘关系,证明了RAPD适用于苏铁属种间亲缘关系分析。RAPD聚类分析结合形态学研究表明:海南苏铁、台湾苏铁、广东苏铁、滇南苏铁和仙湖苏铁之间的亲缘关系较远,支持各自成为独立的种。     

Field confirmation of a mechanism causing synergism between Bacillus thuringiensis and the gypsy moth parasitoid, Apanteles melanoscelus

In 16-ha plots aerially sprayed with single and double applications of Bacillus thuringiensis, percentage parasitism by A. melanoscelus and the number of A. melanoscelus cocoons under burlap strips were higher than in comparable untreated plots in the same area. Strong correlations occurred between percentage parasitism and caterpillar size, with plots having the smallest caterpillars being the most heavily parasitized. However, these parameters were also negatively correlated with number of caterpillars per plot. The increased numbers of parasitoid progeny, i.e., cocoons, found in treated plots showed that corresponding increases in percentage parasitism could not be due simply to improved parasitoid: host ratios. Evidence strongly suggests that the retarding effect of B. thuringiensis infection kept gypsy moth larvae small enough in the treated plots to permit A. melanoscelus females to parasitize relatively large numbers of caterpillars.     

Assessment of genetic fidelity of micropropagated date palm (Phoenix dactylifera L.) plants by RAPD and ISSR markers assay

RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter-Simple Sequence Repeats) markers assay were employed to validate the genetic stability of date palm (Phoenix dactylifera L.) plants multiplied through somatic embryogenesis with upto forty two in vitro subcultures. Out of the 160 RAPD and 21 ISSR primers screened, 30 RAPD and 12 ISSR primers produced a total of 347 (246 RAPDs + 101 ISSRs) clear, distinct and reproducible amplicons, which were monomorphic across all micropropagated plants (27) studied. Thus, a total 8592 bands (number of plants analysed x number of amplicons with all the primers) were generated which exhibited homogeneous banding patterns with both RAPD and ISSR markers. These results indicate that the micropropagation protocol developed by us for rapid in vitro multiplication is appropriate and suitable for clonal propagation of date palm and corroborated the fact that somatic embryogenesis can also be used as one of the safest modes for production of true-to-type plants.     

Improved rearing and release procedures forCotesia melanoscela (Hym.: Braconidae), an early season parasitoid ofLymantria dispar (Lep.: Lymantriidae)

Rearing and release procedures forCotesia melanoscela (Ratzeburg) (Hymenoptera: Braconidae), an early-season parasitoid ofLymantria dispar (L.), were evaluated in 1995 and 1996 at the Beltsville Agricultural Research Center, Beltsville, Maryland, USA. Mass rearing procedures were developed during the winter/spring of 1995/1996 to produce diapausingC. melanoscela cocoons for experimentation. Program efficiency as measured by percent adult emergence was estimated to be 84% with a measured sex ratio of 48:52 M:F, at an estimated cost per 1 000 cocoons of U.S. $5.26 for materials and a labor requirement of approximately 6 hours. A host larvae:female parasitoid ratio of 100:1 was found to be most efficient in terms of number of parasitoid cocoons produced per female parasitoid, but a ratio of 100:2 was most efficient in terms of number of cocoons per rearing cup. OverwinteringC. melanoscela cocoons under natural conditions for 25 weeks (November placements) before expectedL. dispar egg hatch (mid-April) resulted in excellent synchrony with host egg hatch. Alternatively, holdingC. melanoscela cocoons in cold storage for 23 or more weeks at 5°C resulted in adult parasitoid emergence beginning 10 days after removal from cold storage, and proper timing of removal would result in similar synchrony of adult emergence with host egg hatch. These alternative release strategies give managers increased program flexibility.     

FORECASTING GYPSY MOTH DEFOLIATION WITH A GEOGRAPHICAL INFORMATION SYSTEM

Abstract In this study, we developed a series of logistic regression models for forecasting gypsy moth defoliation. The models were parameterized by using data collected over a small scaled 100m× 100m gridarea; the independent variables included egg mass density, male moth trap, previous year defoliation, and distance to the nearest cell which was defoliated in the previous year. We simulated the decision-making by using these models; this method essentially simulated the application of current gypsy moth management decision-making. The results indicated that these models can be more reliably applied to actual management decision.     

中国17个优良玉米自交系的分子标记杂合性及其与杂交种性状的关系研究

在玉米单交种育种中 ,鉴定高产杂交种和具有优良特性的自交系是一个重要的问题。研究以 1 7个优良玉米自交系为亲本 ,按照双列杂交配组合 ,利用 RAPD技术分析了 1 7个自交系的多态性以及 RAPD标记与 9个重要农艺性状 (包括产量 )的关系。基于 RAPD标记计算的相似系数聚类将 1 7个自交系分为 5个类群 ,经分析与系谱亲缘关系基本一致。杂交种性状及其特殊配合力与亲本间的遗传距离是高度相关的 ,与聚类前比较 ,聚类后平均遗传距离与平均产量、平均特殊配合力的相关系数显著提高 ,类间平均产量高于类内平均产量。RAPD技术可揭示优良玉米自交系的系谱亲缘关系 ,将自交系划分成不同的类群 ,从而为选择类间自交系杂交 ,进行亲本选配和分子标记辅助育种提供一种方法。     

香蕉炭疽菌菌株亲缘关系的RAPD分析*

应用筛选的8个Operon随机引物对供试菌株的DNA进行扩增,所产生的RAPD图谱揭示了香蕉炭疽菌具有丰富的种内遗传多样性。UPGMA聚类分析的结果表明：26个香蕉炭疽菌被分为两个与地理来源相关的RAPD聚类群,它们分别由广东菌株和海南菌株为主组成。在两个香蕉菌群的外侧,两个胶胞炭疽菌菌株聚为一个小的外群。这些结果表明香蕉炭疽菌存在有与地域相关的种下类群分化。     

香蕉炭疽菌菌株亲缘关系的RAPD分析

应用筛选的8个Operon随机引物对供试菌株的DNA进行扩增,所产生的RAPD图谱揭示了香蕉炭疽菌具有丰富的种内遗传多样性。UPGMA聚类分析的结果表明：26个香蕉炭疽菌被分为两个与地理来源相关的RAPD聚类群,它们分别由广东菌株和海南菌株为主组成。在两个香蕉菌群的外侧,两个胶胞炭疽菌菌株聚为一个小的外群。这些结果表明香蕉炭疽菌存在有与地域相关的种下类群分化。     

用地理信息系统实现舞毒蛾危害的预测(英文)

本文提出了预测舞毒蛾危害造成树木失叶的逻辑斯蒂回归模型。数据调查是以100m×100m为一格的小尺度下进行的,回归自变量是卵密度、雄蛾密度、前一年的失叶情况和到前一年最近的失叶格的距离,预测结果是一个失叶概率的分布图。我们用文中模型模拟了目前舞毒蛾综合管理中的防治决策过程,结果表明本文提出的模型在应用于实际的管理决策中是很合适的。     

SDS-PAGE、同工酶等电聚焦和RAPD标记鉴定两系杂交水稻F1种子纯度对比研究

目的：探讨利用种子贮藏蛋白SDS—PAGE电泳、酯酶同工酶超薄等电聚焦电泳和RAPD分子标记鉴定五个两系杂交水稻组合F_1种子纯度的可行性。方法和结果：利用SDS—PAGE电泳技术未能找到所试五个组合各自的父本特征蛋白质带;利用酯酶同工酶超薄等电聚焦电泳技术和RAPD分子标记可找到所试五个组合的父母本特征酶带和RAPD标记,但酯酶同工酶的多态性同时也受种子萌发时间的影响。酯酶同工酶超薄等电聚焦电泳和RAPD分子标记可用于所试五个两系杂交水稻组合F_1种子纯度的鉴定。     

RAPD技术在微生物生物多样鉴定中的应用

RAPD（随机放大多态性DNA）是1种新的DNA分子标记技术。与RFLp、AFLP及ARDRA相比,RAPD具有可在一次试验中同时观察到大量的DNA多态性片段,方法更具简单、敏感、花费少等优点。阐述了RAPD的原理方法,及目前在微生物分类鉴定研究中的应用,并分析了RAPD技术在共生固氮放线菌Frankia分类鉴定及系统发育研究中的应用前景。     

RAPD analysis of genetic variation between a group of rose cultivars and selected wild rose species

The genetic variability based on random-amplified polymorphic DNA markers was analysed among 10 cultivated rose varieties and 9 wild species from three different series of the genus Rosa. Using 13 different RAPD primers, 104 polymorphic DNA fragments with a high potential to differentiate rose genotypes could be produced. A dendrogram displaying the relative genetic similarities among the genotypes shows the existence of large genetic diversity among the cultivated roses as compared to the wild species. Furthermore, the main clusters found here are in agreement with known pedigrees and the classical taxonomy. However, the relationships between cultivated roses as inferred by RAPD markers do not correlate with the classical rose classification system. From the present data it is concluded that cultivated roses display a high level of genetic variability despite the fact that single morphological and physiological characters may be less polymorphic within rose groups. This contrasts with the widely accepted opinion of a lack of genetic variability in roses. This is also in accordance with the reported history of rose breeding which makes it highly probable that rose genomes comprise mosaics of different species genomes. As a consequence, it may be possible to utilize the high genetic variability of all genetic traits not under actual selection by breeders for future breeding programmes.     

18种绣线菊分子身份证的构建和SCAR标记转化

取北方常见的18种绣线菊,用200条RAPD随机引物进行扩增,选出10条引物扩增出的清晰稳定、重复性好的图谱进行数据分析。10条引物共检测出51个位点。对凝胶电泳得到的谱带统计结果并赋值,用IDAnalysis 1.0软件进行数据分析的结果表明,仅需6条引物对应的7个位点可将18种绣线菊完全区分。在RAPD扩增过程中,找到三裂绣线菊的特异标记SL700,并将此标记转化成SCAR标记,这一特异标记在分子水平可将三裂绣线菊与其他17种绣线菊区分开来。     

NABIC marker database: A molecular markers information network of agricultural crops

In 2013, National Agricultural Biotechnology Information Center (NABIC) reconstructs a molecular marker database for useful genetic resources. The web-based marker database consists of three major functional categories: map viewer, RSN marker and gene annotation. It provides 7250 marker locations, 3301 RSN marker property, 3280 molecular marker annotation information in agricultural plants. The individual molecular marker provides information such as marker name, expressed sequence tag number, gene definition and general marker information. This updated marker-based database provides useful information through a user-friendly web interface that assisted in tracing any new structures of the chromosomes and gene positional functions using specific molecular markers.

Availability

The database is available for free at http://nabic.rda.go.kr/gere/rice/molecularMarkers/     

大豆分子标记在RIL群体中的偏分离分析

利用栽培大豆与半野生大豆杂交得到的Ｆ８代重组自交系,对２３８个分子标记的偏分离现象进行了分析。结果表明,２９．４％的位点出现偏分离,并且有偏向母本“长农４号”的趋势。