DNA分子标记技术在花生品种鉴定上的应用研究

利用10个中国花生品种,评估了AFL,SRAP,SSR和STS技术在区分相近来源花生品种上的应用效果。结果表明,AFLP和SRAP标记虽然可以揭示这些花生品种的遗传多样性,但在本研究采用的引物对条件下,单独使用任何一对引物不足以区分全部品种,提示我们需要考虑多对引物的配合,或者需要尝试更多的限制酶或引物对;从103对SSR或STS引物中筛选出9对引物,即Lec_1,Ah4_26,Ah4_4,SsS14,SHPAL_1,PG_71,PG_43,PM_36,PG_22,对所采用的10个花生品种的区分率均为100%。鉴于花生栽培种形态学、细胞学和生化标记贫乏,这9对高辨别力的SSR和STS引物对花生遗传研究和品种鉴定工作具有重要价值。     

花生ISSR分子标记技术初步研究

以花生为研究对象,初步建立起花生的ISSR分子标记技术体系,包括DNA提取、DNA模板浓度、引物浓度、退火温度、循环次数和引物筛选等.结果表明:从48条引物中选择4个条带清晰的引物对7个品种进行扩增,共检测出15条带,其中多态性带为12条,多态性比例为80％,能把7个花生品种区分开采.本试验为花生品种鉴定和新品种选育提供理论依据.     

氮肥运筹方式对豫单2002产量及品质的影响

【研究目的】该研究旨在为豫单2002的推广应用提供施肥技术支撑,为高产高蛋白夏玉米氮肥合理运筹肥提供理论与技术依据。【方法】在豫北潮土区高产田块上,采用田间试验,设N0（不施氮肥）、N1（攻杆肥100%）、N2（攻穗肥100%）、N3（攻杆肥40%+攻穗肥60%）,N4（攻杆肥30%+攻穗肥50%+攻粒肥20%）五个处理开展研究。【结果】结果表明：氮肥不同运筹方式增产效果差异显著,其中N3处理增产效果最好,其次是N1处理和N4处理,N2处理增产效果最小;分次施肥有于提高籽粒粗蛋白含量,且随着施肥次数的增加,籽粒粗蛋白含量呈上升趋势;分次施肥亦有利于提高籽粒蛋白质产量,不同处理以N3最佳,其次为N4;氮肥不同运筹方式肥料利用率差异显著,其中N3处理最高,其次为N4和N1处理,而N2最低。【结论】N3处理由于有效地促进了植株对氮素的吸收利用,增加显著地产量和提高了籽粒蛋白质产量,一定程度上提高了籽粒蛋白质含量,因而为最佳氮肥运筹方式。     

分子标记技术在咖啡育种上的应用

主要介绍了AFLP和RAPD 2种分子标记技术以及2种分子标记的优缺点。同时,着重介绍了2种分子标记在咖啡育种上的应用成果。在最后还列举了2个其他的分子标记在咖啡育种上的应用。     

生物技术在蚕豆上的应用及前景

分子标记反映了DNA分子的多态性,可以作为研究生物遗传变异和进化关系的重要手段。介绍了常用分子标记技术的原理、方法、特点及其在蚕豆上的应用现状,同时展望了分子标记技术在蚕豆上的应用前景。     

利用AFLP分子标记预测水稻杂种优势

杂种优势是生物界普遍存在的现象,通过测定两个亲本之间的遗传距离可进而预测水稻杂种优势.迄今已提出了许多预测方法,从表型预测、生化预测,到现在的分子标记预测,DNA分子标记的发展为杂种优势预测提供了新的手段.Lee等[1]和Smith等[2]研究发现RFLP遗传距离与F1杂种优势的表现存在高度的相关性,可用于杂种优势的预测.     

SSR分子标记技术及其在构建玉米DNA指纹库上的应用

SSR分子标记技术是在PCR基础上发展起来的一种DNA多态性检测技术,已开始应用于植物基因组研究的各个领域。本文概述了SSR标记的原理、类型、功能及其引物的来源和利用SSR构建玉米DNA指纹库的进展以及玉米DNA指纹库在品种鉴定、品种保护及品种监测等方面的应用。     

目标起始密码子多态性(SCoT)分子标记技术在花生属中的应用

花生属分子标记领域的研究远落后于其他物种,而栽培种花生因其遗传基础狭窄,用大多数分子标记技术都难以检测到丰富的分子标记,因此限制了花生属野生种在改良花生栽培种方面的利用以及建立花生分子标记辅助育种技术体系。本文分别对花生属4个区组的16份种质资源和8份花生栽培种资源采用与功能基因相关的SCoT分子标记技术研究了花生属种间和栽培种内遗传多样性和亲缘关系。23条SCoT引物在花生属试材基因组中的扩增位点共194个,其中多态性位点130个,多态性达67.01%,通过聚类分析研究了它们之间的亲缘关系;在栽培种内筛选出19条多态性引物,在8份试材基因组中扩增位点198个,其中多态性位点67个,多态性为33.84%,表明SCoT分子标记技术能在花生栽培种内检测出一定程度的DNA多态性。     

分子标记在青稞中的应用现状及前景

分子标记反映了DNA分子的多态性,可以作为研究生物遗传变异和进化关系的重要手段.本文介绍了常用分子标记技术的原理、方法、特点及其在青稞上的应用现状,同时展望了分子标记技术在青稞上的应用前景.     

RAPD分子标记技术在甘蔗育种上的应用

PAPD（随机扩增多态性DNA）是一项新兴的分子标记技术。该技术与其它分子标记技术如RFLP、DNA指纹图谱、SSCP等相比具有如下特点：1）对受试基因组无特定的要求，无需知道模板DNA序列的信息；2）不需要使用放射性标记；3）仅需毫微克数量级的DNA样品，一般1次扩增只需10一50ng的DNA；4）扩增引物随机设定，可从许多物种中检测出大量的多态DNA；5）检测灵敏度高，操作简单快捷，适合大量样本的快速分析。因而，RAPD技术正越来越广泛地应用于分子系统学研究、品种鉴定、基因组研究、遗传连锁图谱的构建、基因定位等研究领域。     

分子标记在花生育种中的应用

简述了RFLP、ARPD、AFLP、SSR分子标记在花生育种中的应用,并提出了存在问题及解决方法。为了尽快将分子标记用于花生育种,应提高分子标记的稳定性,降低所需费用,提高自动化操作程度,积极做好中国花生种质资源的分子标记工作,以及与高产优质高抗有关的新基因鉴定利用及其他相关的标记和定位。     

花生分子育种研究进展

随着高通量测序技术、基因组学分析技术和分子生物学技术的发展,分子育种已成为花生育种的重要手段之一。在新一代高通量测序技术的影响下,大量的花生功能基因和分子标记被挖掘出来,遗传连锁图谱更加精细化,强化了分子标记与常规育种的有机结合,促进了花生转基因技术发展。本文对国内外花生分子育种的研究进展进行综述,并对花生分子育种的主要问题和发展前景进行了讨论。     

玉米分子遗传框架图谱构建

以48-2×5003的166个F2单株为作图群体,利用135个RFLP探针和131对SSR引物对亲本48-2、5003之间的多态性进行了检测,筛选出109个RFLP多态性探针和81对SSR多态性引物用于F2群体分析,利用上述109个RFLP标记和81个SSR标记,构建了具190个RFLP、SSR标记199个标记位点的玉米分子遗传图谱,覆盖整个基因组2984.1 cM,标记间平均间     

Molecular markers and doubled haploids in European plant breeding programmes

The breeding companies and laboratories involved in this article cover a wide range of crops grown in the temperate climate zone: small grain cereals, oilseed crops, forage crops, turf, vegetables and potato. Speed and efficiency are becoming increasingly important in variety breeding and doubled haploids (DH) and genetic markers are important biotechnological tools to accelerate materials to market. Collaborative research between universities, research institutions and breeding companies has resulted in the routine use of DH technology and molecular markers in practical breeding of barley, wheat and rapeseed. DH populations have been established not only for barley, wheat and rapeseed, but for rye, oat and triticale, where DH technology is less developed. A driver here is the value of the crop e.g. although wheat is less responsive to DH production the value of the end product makes the effort worthwhile. Simple and rapid DNA extraction methods used in high-throughput marker assisted selection (MAS) systems are essential for routine use of markers. MAS is used both to monitor the presence of genes of interest and also to monitor the genetic background. DH technology in forage, turf and vegetables is still in progress and the practical use of markers in all crops is limited by access to trait linked markers. Collaboration and technology transfer with universities, research institutions and breeding companies is essential for the improvement of both DH protocols in recalcitrant crops and marker technology in all crops.     

Molecular Marker based Genetic Diversity Analysis in Rice (Oryza sativa L.) using RAPD and SSR markers

The availability of an array of molecular marker systems allowed comparing the efficiency of two of these marker systems to estimate the relationships among various taxa. The objective of this study was to assess the genetic diversity among 40 cultivated varieties and five wild relatives of rice, Oryza sativa L. involving simple sequence repeat (SSR) randomly amplified polymorphic DNA (RAPD) markers. The accessions were evaluated for polymorphisms after amplification with 36 decamer primers and 38 SSR primer pairs. A total of 499 RAPD markers were produced among the 40 cultivated varieties and five wild relatives with a polymorphism percentage of 90.0. Out of 38 SSR primer pairs used, only one locus viz., RM115 was monomorphic. The average Polymorphism Information Content (PIC) value was 0.578 and it ranged from a low of zero (RM 115) to a high of 0.890 (RM 202). The Mantel matrix correspondence test was used to compare the similarity matrices and the correlation coefficient was 0. 582. The test indicated that clusters produced based on RAPD and SSR markers were not conserved since matrix correlation value was 0.582 as against the minimum required value of 0.800. The two marker systems contrasted most notably in pair-by-pair comparisons of relationships. SSR analysis resulted in a more definitive separation of clusters of genotypes indicating a higher level of efficiency of SSR markers for the accurate determination of relationships between accessions that are too close to be accurately differentiated by RAPD markers. This revised version was published online in July 2006 with corrections to the Cover Date.     

Applications and impact of molecular markers on evolutionary and diversity studies in the genus Allium

Allium is a large variable genus containing ≅ 700 species, among them the economically important crops bulb onion, garlic and leek and many other crop species of minor or local importance. As the methods became available and more refined, systematic studies using biochemical and molecular markers were published to complement and offset the growing amount of data based on morphological, anatomical and other traditional approaches. Using isozymes and later restriction fragment length polymorphisms (RFLPs) and random amplified polymorphic DNAs (RAPDs), phylogenetic problems were addressed at all systematic levels of the genus and the interrelationships among crop species and their close relatives. The analysis of hybrid plants is another important application of molecular markers. Markers allow verification of the hybrid's cytoplasm, the composition of the nuclear genome, and investigation of introgression in backcrossing experiments.     

基于油楠(Sindora glabra)转录组测序的SSR分子标记的开发

油楠(Sindora glabra)作为极具开发潜力的能源植物,在分子水平上的研究仅局限于ISSR分子标记。本研究利用Trinity软件对油楠转录组数据进行组装,共得到总长为48307806 bp的283998条Unigenes。利用MISA软件寻找到97443个含有SSR的重复基元。SSR位点中主要重复序列为单核苷酸和二核苷酸,分别占总SSR位点的67.68%和22.56%,其次是三核苷酸,占8.54%。利用Primer 3.0设计引物,并从中随机选择200对引物进行PCR扩增,其中有13对扩增出清晰条带,多态性高,重复性好。油楠SSR分子标记的开发对于研究其遗传多样性、重要性状基因定位及分子标记辅助选择育种等起到重要的推动作用。     

Verification of the parthenogenetic capability of unreduced eggs in an alfalfa mutant by a progeny test based on morphological and molecular markers

Apomixis involves the parthenogenetic development of apomeiotic eggs. It has the potential of cloning plants through seed, and thus furnishes a unique opportunity in breeding of allogamous sexual species, such as alfalfa, for developing superior cultivars with permanently fixed heterosis. Apomixis as a whole has not been detected in the genus Medicago, but components of apomixis have been reported. The formation of unreduced eggs through diplosporic meiosis was documented in a diploid mutant of M. saliva ssp. falcata (L.) Arcang., named PG-F9. Since in facultative apomictic species non-reductional meiosis and parthenogenesis could be tightly associated processes, a progeny test based on morphological trait and molecular marker evaluation was carried out to verify the occurrence of parthenogenesis in PG-F9. Morphological traits such as leaf shape, stipule form, stem pigmentation and flower colour were shown to be effective in the preliminary screening of progenies and most of the plants were classified as non-maternal (i.e. from sexual reproduction). Molecular investigations by means of random amplified polymorphic DNA (RAPD) fingerprint and heterozygous restriction fragment length polymorphism (RFLP) loci detection conducted on the progenies classified morphologically as maternal allowed two plants, molecularly similar but not identical to PG-F9, to be discovered. Owing to the high number of molecular markers conserved as in the mother plant, and because of the great discriminating efficiency of the primers and probes used, these progeny plants could most likely be generated through parthenogenesis of diplosporic eggs. In fact, the extraordinary preservation of maternal morphological traits and genomic loci over one generation may be explained only if apomictic reproductive events rarely took place in PG-F9.     

Assessment of genetic variability in grapefruits (Citrus paradisi Macf.) and pummelos (C. maxima (Burm.) Merr.) using RAPD and SSR markers

The genetic variability of 38 grapefruit (Citrus paradisi Macf.) and three pummelos (C. maxima (Burm.) Merr..) accessions was evaluated using RAPD, and single sequence repeat (SSR) analyses. Approximately49% of the 198 RAPD were polymorphic, and 4.6 alleles per SSR loci were identified. PIC values changed from 0.093 to 0.450. A UPGMA phenetic tree was constructed and two main grapefruit groups were identified. The grapefruit accessions `do Cabo' and `Siamesa-Filipinas'clustered very close to the pummelos in Group A. The Group B consisted of three sub-groups, which comprised all of the other grapefruit accessions. The majority of grapefruit accessions showed a narrow genetic base suggesting that the observed morphological polymorphism within the group must be associated with somatic mutations, which were not detected by these molecular markers. This revised version was published online in August 2006 with corrections to the Cover Date.     

分子标记在木兰科植物研究中的应用

林木研究中常采用的分子标记技术主要包括RFLP、RAPD、AFLP、SSR及ISSR等。本文综述了这些分子标记技术的原理、优缺点。归纳总结了分子标记在木兰科植物中的应用研究进展:(1)利用RAPD、RFLP、cpDNA基因系列测定(psbA-trnH、atpB-rbcL、matK、ndhF)等分子标记在分子水平上对一些群体、个体进行了亲缘关系和分类研究;(2)利用RAPD、SSR和ISSR标记对一些群体、个体进行了遗传结构和遗传多样性研究;(3)采用DAF和RAPD获得了厚朴的DNA指纹图谱。分子标记在木兰科植物的其它方面的应用还很少。今后,除了继续对上述方面进行深入系统的研究外,还应充分运用分子标记技术,开展木兰科植物的分子遗传图谱、分子标记辅助选择育种、保育生物学等方面的研究。