三系杂交稻亲本随机扩增多态性DNA(RAPD)分析

选用9个随机引物对31份杂交水稻亲本材料进行了RAPD分析, 共检测到60条多态性带。聚类分析结果表明,所有供试材料可以被明确地区分。在9个随机引物中,有8个具有较高的多态性检测能力。以这8个引物为基础,选用任两个引物即可在任一对材料中检测出多态性的频率在96.13%以上,而选用任3个引物则该频率在99.21%以上。这显示了运用RAPD鉴定稻种具有简便、灵敏、高效的优点,在鉴定杂交稻种的实践中有着良好的应用前景。 Abstract:Seven rice sterile lines,12 maintainer lines and 12 restorer lines were analyzed by RAPD with 9 primers.Altogether,118 fragments were generated,of which 60 detected polymorphisms among rice marker.Eight of nine primers can detect high polymorphism.The frequencies of polymorphism in any primers were used,the frequencies would be higher than 99.21%.The eight primers were therefore recommended as candidates for the identification of hybrid rice seeds.     

红莲型杂交稻(红莲2号)及其骨干亲本的RAPD分析与鉴定

利用RAPD技术,从248个随机寡核苷酸引物（10－mer）中筛出18个引物对红莲型杂交稻组合红莲2号及其亲本（T－07A、T－07B、YD6－05）,另6个红莲型胞质不育系的骨干恢复和汕优63及其亲本共14份水稻材料进行分析。共检测到173个多态性标记。聚类分析结果表明：不育系与保持系间因核背景相似,遗传差异很小;杂种（F1）的基因型更倾向于恢复系;恢复系与保持系间遗传距离的相对较大,但各恢复系之间的遗传距离较小。利用这些标记能有效地地区交组合中不育系,保持系、恢复系和杂种（F1）。     

用RAPD分子标志方法研究氧化亚铁硫杆菌遗传多样性

本文对采自7个不同环境的氧化亚铁硫杆菌进行了随机扩增多态性DNA（RAPD）分析,20个引物中筛选出扩增效果较好的4个引物,每个引物能产生1~9条DNA条带。通过4个引物的RAPD分析获得的平均相似性系数表明不同来源的菌之间的相关系数在44%~83%之间。 Abstract:Random amplified polymorphic DNA(RAPD) was used in analyzing the polymorphisms of Thiobacillus ferrooxidans from seven different places.Of the 20 primers,four could generate reproducible RAPD profiles,and each one produced 1~9 bands.The similarity coefficients obtained from profiles generated by four primers among Thiobacillus Ferrooxidans were about 44%~83%.     

BT型细胞质雄性不育水稻及其三系的线粒体DNA研究

用RAPD技术对BT型水稻胞质雄性不育系秀A及其保持系秀B、恢复系湘晴以及杂种F1代的线粒体DNA进行了比较分析。结果表明不育系与其保持系间存在显著差异;不育系与其F1之间mtDNA也存在差异。在引物OPJ－08的扩增产物中,秀A扩增出一条分子量为800bp的多态性片段,在引物OPK－10的扩增产物中,杂种F1扩增出一条分子量为900bp的片段。把这两片段回收、克隆并制备探针,OPJ－08800的Southern杂交结果显示不育系与其F1杂交图谱存在多态性;OPK－10900的Suthern杂交结果显示不育系与其保持系同存在差异。推测这两片段与育性可能有一定的联系。     

粘类小麦雄性不育恢复基因的遗传分析及RAPD标记

利用RAPD分子标记对粘类小麦雄性不育系ms（Kots）-90-110的恢复系Rk5451的恢复基因进行了标记定位。选取具有高恢复力的恢复系康本材料Rk5451和Rk5253为父本与ms（Kots）-90-110杂交．F1代再与保持系90-110回交;以90-110／／ms（Kots）-90-110／Rk5451的BC1F1代分离群体为研究对象．利用分离群体分组分析法（Bulked Segregant Analysis．BSA）．以350个随机引物对Rk5451的主效恢复基因进行RAPD分析．筛选到27个可在亲本间扩增出多态性的引物．其中引物S120经多次重复能在亲本间及不育和可育池间扩增出稳定的多态性片段S120-1745。     

利用RAPD标记构建美洲黑杨×欧美分子标记连锁图谱

本文利用RAPD标记和美洲黑杨(Populus deltoides)×欧美杨(P.euramericana)的F1群体,构建了美洲黑杨×欧美杨的分子标记连锁图谱。实验过程中对1040个寡核苷酸随机引物进行了重复筛选,共选出127个引物用于作图群体(包括双亲共92个无性系)的随机扩增,这127个引物产生229个多态基因座,其中符合“拟测交”1∶1分离的有214个。利用多点连锁分析,形成19个连锁群及6个三连体和14个连锁对。由19个连锁群构成的图谱含标记129个,总图距为1914.2cM,覆盖杨树基因组约73.62％。标记间的平均间距为14.84cM。本研究获得了中等密度的美洲黑杨×欧美杨的一个连锁框架。 Abstract:A molecular linkage map was constructed for the parents of a P.deltoides × P.euramericana F1 family based on random amplified polymorphic DNA(RAPD)markers.A set of 1040 random oligonucleotide primers were screened and 127 primers were selected to generate RAPD markers within a sample of 90 F1 progenies.A total of 229 segregating loci were identified.Among the 229 loci,15 loci were found distorted from the normal 1∶1 ratio.Using multiple analysis,the 214 markers formed 19 main Linkage groups (including 129 markers)and b triples and 14 pairs.The resulting Linkage map of Populus deltoides × P.euramericana (including 129 markers)spanned 1914.2cM(73.62％ coverage of genome length)with an average distance of 14.84cM between markers.     

水稻孢子体细胞质雄性不育系线粒体DNA的RAPD分析

利用200个10nt随机引物对水稻孢子体细胞质雄性不育系珍汕97A、保持系珍汕97B、F1代汕优63、恢复系明恢63和另一种孢子体细胞质雄性不育系马协A、保持系马协B、F1代马协63的线粒体DNA进行PCR扩增。在36℃的退火温度下,有132个引物扩增出了清晰的、重复性较好的条带。同一组合内的不育系同保持系、恢复系之间线粒体DNA随机引物扩增产物的多态性明显高于不育系与F1代。不育系与F1代线粒体DNA的扩增产物也存在多态性。一些特异性的差异片段可能与水稻CMS相关。     

利用RAPD标记构建美洲黑杨×欧美分子标记连锁图谱

本文利用RAPD标记和美洲黑杨(Populus deltoides)×欧美杨(P.euramericana)的F1群体,构建了美洲黑杨×欧美杨的分子标记连锁图谱。实验过程中对1040个寡核苷酸随机引物进行了重复筛选,共选出127个引物用于作图群体(包括双亲共92个无性系)的随机扩增,这127个引物产生229个多态基因座,其中符合“拟测交”1∶1分离的有214个。利用多点连锁分析,形成19个连锁群及6个三连体和14个连锁对。由19个连锁群构成的图谱含标记129个,总图距为1914.2cM,覆盖杨树基因组约73.62％。标记间的平均间距为14.84cM。本研究获得了中等密度的美洲黑杨×欧美杨的一个连锁框架。 Abstract:A molecular linkage map was constructed for the parents of a P.deltoides × P.euramericana F1 family based on random amplified polymorphic DNA(RAPD)markers.A set of 1040 random oligonucleotide primers were screened and 127 primers were selected to generate RAPD markers within a sample of 90 F1 progenies.A total of 229 segregating loci were identified.Among the 229 loci,15 loci were found distorted from the normal 1∶1 ratio.Using multiple analysis,the 214 markers formed 19 main Linkage groups (including 129 markers)and b triples and 14 pairs.The resulting Linkage map of Populus deltoides × P.euramericana (including 129 markers)spanned 1914.2cM(73.62％ coverage of genome length)with an average distance of 14.84cM between markers.     

五种品系猪亲缘关系的RAPD分析

应用190个RAPD引物对贵州小型猪,巴马小型猪、西双版纳近交系小耳猪JB和JS亚系、荣昌猪Ⅰ系、长白猪的亲缘关系进行了初步分析,其中39个引物的扩增结果具有非常明显的品系间多态性,将其扩增结果以RAPDistance package Version 1?04程序进行分析,计算相对遗传距离指数1－F,再用NJ法进行聚类分析,构建系统树。结果表明,三种小型猪相互间亲缘关系较近,尤以贵州小型猪与巴马小型猪更近。荣昌猪Ⅰ系和长白猪关系较近,但与小型猪关系较远。 关键词：随机扩增多态DNA;小型猪;亲缘关系 The phylogenetic relationship of five species of pigs including Guizhou miniature pig,Bama pig,Xisuangbanna inbred pig,Rongchang Pig and Landrance was studied by RAPD analysis.Thirty-nine single polymorphic primers were selected out of 190 primers.The amplified fragments of thirty-nine primers were analysed by the RAPDistance package version 1.04 and the phylogenetic tree was constructed using NJ method.The results indicated as the following: three strains of miniature pigs have close phylogenetic relationship and the phylogenetic relationship between Guizhou miniature pig and Bama miniature pig was much closer.Landrance and Rongchang pig have close phylogenetic relationship too,but their phylogenetic relationship is far from the three strains of miniature pigs.     

水稻抗白叶枯病基因Xa-4的PCR标记研究

根据与水稻抗白叶枯病基因Xa-4紧密连锁的分子标记M55的序列设计引物,通过对国际水稻研究所育成的抗白叶枯病近等基因系和基因累加系的叶片DNA、半粒种子提取物及Xa-4基因的杂合体DNA的PCR特异扩增,初步建立了Xa-4的PCR标记体系。进而用该标记体系对我国籼型杂交水稻常用的亲本材料进行分析,揭示出了Xa-4在这些材料中的分布情况。 Abstract　Based on the sequence of a DNA marker tightly linked to the rice bacterial blight（BB） resistance gene Xa-4, two primers were designated and synthesized to develop a PCR marker for the gene. Specific amplified polymorphism analysis was carried out with these primers on a set of BB resistance isogenic lines and pyramided lines developed by IRRI. Two PCR bands were revealed corresponding to lines with dominant Xa-4 and those with the recessive allele, respectively, regardless the lines pyramided with other resistance genes. A hybrid with heterozygous Xa-4 produced both of the two allele PCR pattern. Then, the PCR marker was used to survey a range of hybrid rice germplasm. The results of the germplasm survey will be useful in hybrid rice breeding programs aimed at exploiting Xa-4.     

云南普通野生稻遗传多样性和亲缘关系

野生稻（Oryza rufipogon）是稻属的重要组成部分,具有许多优良性状,是水稻遗传改良的天然基因库。本研究通过对形态学性状的观测,及ISSR和RAPDUPGMA聚类分析,将云南普通野生稻划分为4个类型,即元江类型、景洪紫杆直立型、景洪绿杆直立型和景洪匍匐型。在供试材料中筛选到具有多态性的ISSR和RAPD引物各11个,ISSR引物扩增出多态带113条,多态性条带比率（PPB）为82．26％,RAPD引物共扩增出多态性条带76条,PPB值为76．77％,两种分子标记的分析结果呈极显著正相关（r=0.951）。此外UPGMA聚类结果表明,云南普通野生稻不同类型与其它地区普通野生稻之间的遗传亲缘关系差异明显。     

云南普通野生稻遗传多样性和亲缘关系

野生稻(Oryza rufipogon)是稻属的重要组成部分, 具有许多优良性状, 是水稻遗传改良的天然基因库。本研究通过对形态学性状的观测, 及ISSR和RAPD UPGMA聚类分析, 将云南普通野生稻划分为4个类型, 即元江类型、景洪紫杆直立型、景洪绿杆直立型和景洪匍匐型。在供试材料中筛选到具有多态性的ISSR和RAPD引物各11个, ISSR引物扩增出多态带113条, 多态性条带比率(PPB)为82.26%, RAPD引物共扩增出多态性条带76条, PPB值为76.77%, 两种分子标记的分析结果呈极显著正相关(r = 0.951)。此外UPGMA聚类结果表明, 云南普通野生稻不同类型与其它地区普通野生稻之间的遗传亲缘关系差异明显。     

水稻广亲和基因（WCG）近等基因系的随机引物PCR分析

应用随机引物ＰＣＲ（ＲａｎｄｏｍＰｒｉｍｅｒＰＣＲ）技术分别在水稻广亲和基因（ＷＣＧ）的近等基因系和色素原基因（Ｃ）的分离群体库中寻找与ＷＣＧ和Ｃ基因连锁的分子标记。对于ＷＣＧ近等基因系,在２２６个随机引物中初筛到２２个显示多态性片段的引物。根据理论值计算,在２２个多态性片段中预期有２０个与ＷＣＧ连锁。在这些连锁标记中距ＷＣＧ最近的可达０．５ｃＭ。同样在分离群体库的筛选中有１０个扩增产物与Ｃ基因连锁。     

RAPD和ISSR标记对水稻化感种质资源遗传多态性的分析

运用RAPD和ISSR技术分析水稻化感种质资源的遗传多态性。从供试材料中筛选到具有多态性的RAPD引物12条,ISSR引物7条。RAPD引物共扩增到85条清晰的多态性条带,多态性条带比率为69．4％。ISSR引物共扩增到34条清晰的多态性条带,多态性条带比率为53．0％。对两种标记结果进行UPGMA聚类分析,结果极其类似,呈极显著的正相关(r=0．74)。聚类结果表明,地理位置相近的品种聚为一类。部分具有较强化感作用潜力的水稻品种亲缘关系很近,表明控制其化感作用性状的基因可能是等位的相同基因。而部分化感作用潜力差异显著的水稻品种聚为一类,这是由于人类在长期高产品种的定向选择过程中,水稻化感作用性状不被注意而丢失,遗传基础日益狭窄的原因。     

DNA Polymorphism among Rice Somaclones

Molecular markers were used to detect the influence of high concentrations of 2,4 dichlorophenoxyacetic acid (2,4-D) in the callusing media on DNA variations in regenerated rice plants. Restriction fragment length polymorphism (RFLP), random amplified polymorphic DNA (RAPD) and polymerase chain reaction (PCR) based RFLP analysis were carried out on 12 somaclones of Oryza sativa L. cv. B-370. In vitro culture induced DNA variations were detected in the regenerated plants but the effect of high auxin concentration in the medium could not be revealed. In a second study, fingerprinting of 15 semi-dwarf, high yielding somaclones of B-370 was carried out using RAPD technique. Amplification using 20 random primers produced a total of 167 DNA bands out of which 97 bands were polymorphic. A total of 32 unique DNA bands were detected across all the somaclones and they could be grouped based on their similarity to B-370. RAPD analysis helped to reveal similarity or differences among the somaclones while fingerprinting using additional RAPD markers was not successful.     

Genetic variation in monoploids of diploid potatoes and detection of clone-specific random amplified polymorphic DNA markers

Randomly amplified polymorphic DNA (RAPD) markers have been used to study the genetic variation among androgenetic monoploids of diploid Solanum species. Cluster analysis of pairwise genetic distances was used to construct a genetic relationship among anther donor and anther-derived potato plants. The clustering based on Rogers' distances resembled classifications based on parental origins and hybrid combinations. Six of the 32 RAPD primers used resulted in the selective amplification of DNA fragments which were polymorphic between the two S. phureja parental clones, 1.22 and A95. It should be possible to construct a genetic linkage map, without making crosses, using monoploids derived from a single heterozygous diploid clone and RAPD markers.     

Inheritance of RAPD markers in channel catfish (Ictalurus punctatus), blue catfish (I. furcatus), and their F1, F2 and backcross hybrids

The channel catfish ( Ictalurus punctatus ) has become the most important aquaculture species in the USA. A genetic linkage map in catfish is needed to improve efficiency of breeding by marker-assisted selection (MAS) and for identification of economically important genes such as disease resistance genes. To identify DNA-based genetic polymorphism, the present authors tested 42 randomly amplified polymorphic DNA (RAPD) primers for their utility in identifying genetic polymorphism in catfish. Out of these primers, 22 generated 171 highly reproducible RAPD markers, producing almost eight polymorphic bands per primer. The remaining 20 primers produced an additional 20 polymorphic bands. The RAPD markers were highly reproducible, transmitted to F1 hybrids, and segregated in F2 or backcross progeny in ratios that did not differ from Mendelian expectations. Because the interspecific hybrids of channel catfish and blue catfish are fertile, RAPD markers using the interspecific hybrid system will be useful for rapid construction of genetic linkage maps of catfish and for analysis of important quantitative trait loci.     

Analysis of Genetic Diversity in Napier Grass (Pennisetum purpureum Schum) as Detected by RAPD and ISSR Markers

Randomly Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeat (ISSR) markers were used to detect the DNA polymorphism among thirty Napier grass collections of wide geographical distribution. A total of 20 RAPD and 10 ISSR primers were used in this study. RAPD analysis produced 222 fragments of which, 195 were polymorphic with an average of 9.75 polymorphic fragments per primer. The ten ISSR primers produced a total of 98 fragments out of which 88 were polymorphic accounting for 89.8%. The Mantel test between two similarity matrices of the markers revealed a low correlation (r = 0.33) indicating low correspondence between polymorphism brought out by the two marker systems. The UPGMA clustering of genotypes eventhough was not similar when RAPD and ISSR derived dendrograms were compared, but showed a greater correspondence with geographical identity in both the marker systems employed. This correspondence was also evident when data from both the RAPD and ISSR markers were combined. The implications on collection and breeding of this important forage grass had been discussed.     

Partial molecular characterization of some kiwi fruit cultivars by RAPD markers

Molecular variability among seven cultivars of A. deliciosa var. deliciosa was investigated through RAPD markers. Thirty four decamer primers were screened generating polymorphic patterns of amplified DNA for these cultivars. Twenty one selected primers gave clear and reporducible patterns. A total of 430 bands were produced and 29.37% of them were polymorphic. The patterns distinguished between the cultivars and their analysis established an approach to classification within A. deliciosa var. deliciosa based on RAPD markers. The dendrogram clearly differentiated male from female cultivars. While abbot and allison female cultivars were closely related, bruno and abbot female cultivars showed maximum dissimilarity.