“中国春”小麦不同缺体的RAPD分析

利用ＰＣＲ－９０Ａ型ＤＮＡ扩增仪，对７个中国春小麦缺体核ＤＮＡ进行了扩增。结果显示，中国春小麦缺体核ＤＮＡ的ＲＡＰＤ产物可分为二种类型：（１）所有供试材料均能扩增出的片段，代表了核ＤＮＡ在进化上的保守性。有一个引物检测到这类片段。（２）全部供试材料间存在的差异片段，这类片段是用来基因定位的主要依据。另外，建立中国春小麦缺体系统的ＲＡＰＤ指纹图谱对研究小麦的系统进化也将有重要意义     

桃果实有毛/无毛、白肉/黄肉性状的RAPD分子标记

以桃品种京玉和美味的正反交后代６９株为试材,采用ＲＡＰＤ指纹分析技术,获得了控制桃果实性状有毛／无毛、白肉／黄肉这２个基因的ＲＡＰＤ分子连锁标记,其中多态性扩增ＤＮＡ片段ＯＰＰ２０－２２００与果实有毛／无毛基因的连锁遗传距离为５．０ｃＭ,多态性扩增ＤＮＡ片段ＯＰＵ０３－８５０与果实白肉／黄肉基因的连锁距离为９．６ｃＭ。     

RAPD方法在节瓜种子纯度鉴定中的应用初探

本文以节瓜种子为材料,用８种随机引物对节瓜种子父本,母本及杂种一代的染色体组ＤＮＡ进行ＲＡＰＤ扩增,发现有３条随机引物扩增出ＤＮＡ片段,其中２条随机引物属于“偏父型”;１条随机引物属于“特异型”随机引物,另外５条随机引物没有扩增出ＤＮＡ片段。由于随机引物扩增片段多态性的存在,因此可以使用该方法进行种子纯度鉴定。     

利用RAPD技术鉴定大白菜主栽品种及品种间遗传多样性分析

采用随机引物扩增多态ＤＮＡ（ＲＡＰＤ）技术分析了２１个大白菜主栽品种。用１３个引物共扩增出８７个清晰可重复的ＤＮＡ片段,其中３９条带具有多态性,多态性条带的频率为４４．８％。ＯＰＥ０１是多态性频率最高的引物,它可区分１５个大白菜品种,再与引物ＯＰＨ０３和ＯＰＨ１２配合,即可将２１个大白菜品种区别开。     

棉花枯萎病菌生理小种的分子指纹分析

以我国棉花枯萎菌３个生理小种的２６个代表菌株及国外３ 个不同生理小种菌株进行随机扩增多态性ＤＮＡ（ＲＡＰＤ）分析，共产生了１４０个ＲＡＰＤ分子标记，其中８７８％ 具有多态性。通过聚类分析将供试菌株划分为６个ＲＡＰＤ组，确定了不同小种间的亲缘关系，为确立我国棉花枯萎菌生理小种在国际上的分类地位提供了可靠的分子证据。     

用RAPD方法鉴定杂交辣椒种子纯度

本文用ＲＡＰＤ（随机扩增多态性ＤＮＡ）方法对辣椒种子的父本、母本及杂种一代进行分析研究,筛选出适合辣椒杂种一代种子纯度鉴定的随机引物Ｓ５,并对并进了种子纯度的初步鉴定。ＲＡＰＤ结果和田间鉴定的相比,在误差允许范围内,进一步证明了ＲＡＰＤ方法在蔬菜种子纯度鉴定中应用的可行性。     

利用RAPD标记鉴定大豆种质

本研究以５７个中国大豆祖先吕系及育成品种和１８个美国大豆祖先品系为ＤＮＡ样品来源，通过随机引物ＰＣＲ扩增基因组ＤＮＡ的多态性，探索利用ＲＡＰＤ标记鉴定和相关种质的可能性。研究结果表明，５０个１０摩尔随机引物共扩增可分辩产物２４６个，其中８２．４％的随机引物可产生多态性产物，所扩增产物的５４．４％至少在两个基因毒草境存在差异。上ＰＣＲ扩增产物分别以１和０记录存在与否。扩增产物间的成对比较可产生非相似     

应用RAPD分子标记技术对我国骨干玉米自交系进行类群划分

利用ＲＡＰＤ标记技术,对我国目前２５个主要玉米自交系进行亲缘关系类群划分,本项研究建立了从玉米种子、幼芽以及叶片组织提取微量ＤＮＡ的方法。从ＲＡＰＤ引物试验Ａ至Ｏ共计３００个引物中,筛选出对玉米扩增产的具有多态性的引物４０个,其中具有特别明显多态民生的引物１０个。它们是Ｆ３、Ｏ２０、Ａ１９、Ｍ２、Ｍ６、Ｎ１１、Ｎ１９、Ｃ２和Ｇ１４等。依据１０个引物扩增谱带建立０,１型数据,计算２５个自交系间遗传距     

棉花核DNA的提取及其RAPD分析

ＲＡＰＤ是新发展起来的一种分子标记，近几年在植物遗传育种中得到广泛应用。ＲＡＰＤ分析需要分离高质量的核ＤＮＡ，棉花叶片中多酚类化合物含量高，在ＤＮＡ分离过程中，它们跟核酸、蛋白质发生不可逆反应，从而影响了ＤＮＡ的得率和纯度。我们利用最初提取液中增加甘露糖来阻止多酚化合物跟核ＤＮＡ的共价结合，结果得到了高纯度的ＤＮＡ，该ＤＮＡ适于棉花ＲＭＤ分析，并探索出了棉花ＲＡＰＤ分析的反应适宜条件。     

小麦株高近等基因系的RAPD标记研究

采用ＲＡＰＤ技术，以４个小麦株市基因近等基因系（分别含有rht、Ｒht1、Rht2、Rht3)为材料，对２９６个单一随机引物（１０个核苷酸）进行了筛选。发现２５个引我的扩增产物的近等基因系间表现出特异性，在６次重复试验中，有１８个引物的特异护增片段不能重复，６个引物可以重复２－３资，唯有ＯＰＡＭ０１在全部试验中均能稳定重复，其特异扩增版段ＯＰＡＭ０１１８６０可以作为rht基因的ＲＡＰＤ标记。     

Characterisation of powdery mildew resistant lines derived from crosses between Triticum aestivum and Aegilops speltoides and Ae. mutica

Characterisation of introgressions of powdery mildew resistance into wheat from Aegilops speltoides and Ae. mutica was attempted by the application of cytological analysis of meiotic behaviour and molecular markers, based on isozymes and RAPD (random amplified polymorphic DNA). Cytological results suggest that a chromosome segment of Ae. speltoides has been successfully translocated to a wheat chromosome, while transfers from Ae. mutica appear to consist of whole chromosome substitutions. RAPD analysis detected eight bands in the line Sp1, and six bands in the line Mt1, which may be potential markers for powdery mildew (PM) resistance. In all Mt progenies the Ep-T1 product segregated, suggesting that the resistance gene is associated with the group 7 homoeologue in Ae. mutica. This revised version was published online in August 2006 with corrections to the Cover Date.     

Variation for apparent amylose content of endosperm starch in Triticum durum and Aegilops squarrosa

For breeding to increase amylose content in the starch of hexaploid wheat, either the introduction of genes from Aegilops squarrosa (DD genome) or the development of high-amylose amphiploids between Triticum durum (AABB genome) and Ae. squarrosa is one of the most efficient ways. The high-amylose genes have not been detected in any species of Triticum and Aegilops. Therefore, we assessed the apparent amylose content in endosperm starch of T. durum and Ae. squarrosa accessions supplied from the major gene banks from world wide, and found several accessions with higher amylose content out of 665 T. durum accessions and out of 732 Ae. squarrosa accessions. Genetic analysis indicated that a high amylose trait in Ae. squarrosa is monogenically inherited by a recessive major gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genome-specific markers of tetraploid wheats and their putative diploid progenitor species

The objective of this study was to isolate genome‐specific markers from the genomes of tetraploid wheats and the putative donor diploid species on the basis of random amplified polymorphic DNA analysis followed by cross‐hybridization. Twenty different Triticum and Aegilops species and accessions were analysed by polymerase chain reaction (PCR) using 30 random primers. The polymorphic PCR fragments were then isolated, labelled and used in cross‐hybridization screenings. The hybridization results established that one marker was specific to the Ae. speltoides S genome, two to the A genome, one to the B genome and five to the G genomes of polyploid species (and to the genomes of the corresponding progenitor species). Four markers were identified that were specific to both the B and G genomes. Analysis of the Triticum and Aegilops species and accessions supported the notion that Ae. speltoides is more closely related to the B and G genomes of polyploid wheat species than were other members of the Sitopsis section. The data also indicated that the B and G genomes had originated from different accessions of Ae. speltoides.     

Identification of a new family of tandem repeats in Triticeae genomes

A new family of cereal tandem repeats was isolated, characterised and designated as spelt-1. The family of repeats comprises about 2% of the Aegilops speltoides genome; however, its content differs considerably in the genomes of various Triticeae species. Copy number of the constituent sequence, relative to Ae. speltoides, proved to be 40-60 times reduced in the genomes of tetraploid wheats, 400-fold reduced in the genome of Triticum monococcum, and 1200-2400 times in the genomes of the other 19 Triticeae species studied. Drastic difference in the copy number and homology extent of the spelt-1 family sequences between Ae. speltoides and other diploid species allows the utilisation of these sequences as species-specific telomeric markers for Ae. speltoides, provided stringent hybridisation conditions apply. RFLP (restriction fragment length polymorphisms) analysis of spelt-1 reveals polymorphism between the above species. This study of spelt-1 organisation in different Triticum species provided further substantiation of the polyphyletic origin of the B genome of polyploid wheat. This revised version was published online in August 2006 with corrections to the Cover Date.     

Tan-Spot Resistance Screening of Aegilops Species

Two hundred and twelve accessions of 8 diploid and 10 polyploid species of Aegilops were evaluated for resistance to tan-spot disease of wheat, caused by Pyrenophora tritici-repentis (Died.) Drechs., under greenhouse conditions. One or more accessions of Ae. bicornis, Ae. biuncialis, Ae. Crassa, Ae. columnaris, Ae. cylindrica, Ae. speltoides, Ae. squarrosa. Ae. triaristata. Ae. triuncialis, and Ae. Ovata showed resistance following a 24-hour post-inoculation wet period. With a 36-hour wet period, diploids became only slightly or moderately susceptible and resistant polyploids became susceptible. A 48-hour wet period resulted in still greater susceptibility of both diploid and polyploid species.     

利用比较基因组学开发山羊草属InDel分子标记

为开发和利用小麦野生近缘种的有益基因, 采用比较基因组学方法, 通过拟斯卑尔脱山羊草EST(expressed sequence tag)与小麦UniGene序列的比对分析, 发现山羊草插入/缺失(InDel)位点137个, 在这些位点两端序列设计引物24对, 通过在15个小麦野生近缘属种基因组DNA的扩增分析, 发现11对引物具多态性, 可以作为InDel标记。这些包含突变位点的基因涉及亚细胞定位、蛋白质结合与催化以及代谢等过程。     

Phylogenetic relationships as in Ocimum revealed by RAPD markers

Genetic relationships were examined among thirty germplasm accessions belonging to five Ocimum species using RAPD markers. A very high degree of polymorphism (98.20%) was observed. UPGMA cluster analysis of genetic similarity indices grouped all the accessions into two major clusters corresponding to previously reported botanical sections. Intra-clustering within the two clusters precisely grouped the accessions belonging to one species in one sub-cluster as expected from their genetic background. Our results show that RAPD technique is a sensitive, precise and efficient tool for genomic analysis in Ocimum species, that may be useful in future studies, by assigning new unclassified germplasm accessions to specific taxonomic groups and reclassifying previously classified accessions of other Ocimum species by traditional criteria on a more objective basis. This revised version was published online in August 2006 with corrections to the Cover Date.     

TaMyb2-Ⅱ基因在普通小麦(Triticum aestivum L.)及其近缘种中的单核苷酸多态性分析

以39份抗旱性不同的普通小麦、5份A基因组材料、4份拟斯卑尔脱山羊草（Aegilops speltoides）、6份粗山羊草（Aegilops tauschii）和2份四倍体小麦,分析TaMyb2基因的核苷酸序列长度多态性和单核苷酸多态性,及其与抗旱性的关系。结果发现,TaMyb2在A基因组材料中无目标片段扩增,在其他材料中检测到Ⅰ、Ⅱ、Ⅲ 3种类型序列。经详细分析,TaMyb2-Ⅱ序列长1 606 bp,在供试材料77 088 bp的核苷酸序列中包括34个单核苷酸变异,其中26个SNP,8个InDel,二者出现的频率分别为1/2 965 bp和1/9 636 bp,编码区π值（0.00055）小于非编码区的π值（0.00185）, 说明编码区的遗传变异小于非编码区的遗传变异。从SNP水平上分析,发现普通小麦与其D基因组供体种粗山羊草及四倍体小麦的亲缘关系较近,与B基因组供体种拟斯卑尔脱山羊草的亲缘关系较远。48份材料的TaMyb2-Ⅱ序列共分为18个单倍型（haplotype）,其中haplotype 2、3、5、6、8、9均为旱地栽培的普通小麦品种,说明普通小麦TaMyb2-Ⅱ的这几个haplotype结构可能与抗旱性有关。     

Genetic linkage maps of Citrus sunki Hort. ex. Tan. and Poncirus trifoliata (L.) Raf. and mapping of citrus tristeza virus resistance gene

The RAPD technique was used to identify genetic relationships in 19 accessions, including six species of the genus Chenopodium. A dendrogram was constructed using UPGMA from 399 DNA markers. The molecular data clustered species and accessions into five different groups. Group 1 with three cultivated varieties of C. nuttalliae, Group 2 included eight cultivars and two wild varieties of C. quinoa, Group 3 with C. berlandieri and C. album, Group 4 with two accessions of C. pallidicaule, and Group 5 with 2 accessions of C. ambrosioides. The polymorphic patterns generated by RAPD profiles showed different degrees of genetic relationship among the species studied. A low level of intraspecific variation was found within the accessions of C. quinoa, C. nuttalliae, and C. pallidicaule. The RAPD markers were found to be a useful tool for detecting genetic variation within the genus Chenopodium.     

粗山羊草抗小麦白粉病基因遗传多样性的研究

粗山羊草(Ae.tauschii(Coss.)Schmal.)是普通小麦(T.aestivum L.)抗性改良的宝贵遗传资源。本研究对来自伊朗、前苏联等8个国家和地区的78份粗山羊草进行了小麦白粉病(powderymildew)混合菌种苗期接种鉴定,表现免疫或近免疫的有45份,占57.69％;用一套白粉病菌菌株(共16个)对原产地不同的11份粗山羊草分别进行苗期接种鉴定,结果表明除Y168表现感染外,其它10份材料表现出各自不同的抗性反应,只有来自伊朗的Y219、Y221、Y225和来自的苏联的Ae37能抗所有16个菌株,而没有感染的毒性菌株,说明它们可能含有新的不同于15个已知Pm基因的抗性基因。利用完全双列杂交对原产地不同的5份粗山羊草进行了苗期抗白粉病基因的遗传分析,出现3种不同的抗性基因类型,来自伊朗的3份材料表现单显性(PmA)或双显性(PmA、PmB)2种类型,而来自前苏联的2份材料表现出不同于前两种类型的另一种单显性(PmB)类型。从而为增加普通小麦抗白粉病基因的遗传多样性奠定了基础。