分子标记在玉米遗传育种中的应用及展望

分子标记技术的快速发展,为分子标记辅助选择育种提供了便利,有利于加快育种进程。本文概述了各种常用分子标记RFLP、RAPD、AFLP、SSR、SNP的原理;以及近年来分子标记技术在中国玉米遗传多样性、杂种优势及杂种产量的预测、目标基因定位、指纹图谱构建和杂交种子纯度鉴定等研究方面的应用;并简单提出了分子标记技术在未来玉米育种研究中的应用前景和发展方向。     

DNA分子标记在洋葱遗传育种研究中的应用

DNA分子标记技术的出现大大提高了遗传分析的准确性和选育品种的有效性,在植物遗传育种领域越来越受到重视.洋葱属于二年生植物,一个世代需要三年,具有育种年限长的问题.利用分子标记辅助选择是缩短洋葱育种年限,加快育种进程极其有效的途径.本文综述了DNA分子标记技术在洋葱遗传图谱构建、遗传多样性分析、种质鉴定、鉴定洋葱细胞质类型、重要性状的分子标记辅助选择和数量性状的基因定位等方面的应用,讨论了洋葱遗传育种中分子标记技术的现状及存在的问题,并对洋葱分子标记辅助育种的应用前景进行了展望.指出洋葱高密度的遗传图谱还有待于建成,对于数量性状标记的鉴定,还有待于进一步开发、快速和准确定位的QTL,关于洋葱抗病基因的标记需要深入研究.     

SSR分子标记在小麦遗传育种中应用与研究进展

综述简单重复序列标记(SSR)的特点及优点,重点介绍了SSR分子标记在小麦遗传育种中的应用,即利用SSR分子标记技术基因标记与定位、遗传图谱构建、外源染色体鉴定与标记、种质资源鉴定和遗传多样性研究、辅助育种的研究进展。     

ISSR分子标记技术在植物种质资源研究中的应用

ISSR是在SSR基础上发展起来的一种分子标记技术,兼具SSR、RAPD、RFLP、AFLP等分子标记的优点.本文综述了ISSR分子标记技术在种质资源鉴定和指纹图谱构建、遗传多样性和亲缘关系、基因定位和分子标记辅助选择等方面的研究进展.     

分子标记技术在花生上的应用研究

主要介绍了RFLP、RAPD、SSR、AFLP等四种分子标记技术的原理、方法、特点及其在花生上的应用现状：①RAPD、SSR、AFLP都可以分析花生品种的多样性;②RAPD技术可以对杂交后代进行鉴定;③利用RFLP、RAPD技术绘制花生指纹图谱;④利用分子标记技术研究花生根瘤菌;⑤基于花生的RFLP、RAPD技术的改进。同时,展望了分子标记技术在花生上的应用前景,以期对花生的分子标记育种有所帮助。     

水稻种质资源的分子鉴定和育种利用

水稻种质资源包括栽培稻、野生稻及育种材料等各种类型,是水稻新品种选育、生物技术研究以及农业生产的物质基础。中国具有丰富的水稻种质资源,但其利用率却不高,原因之一就在于传统方法不能够对其进行准确的研究评价。理想的分子标记具有稳定性高、重复性好、检测简单快速和共显性遗传等优点。本文综述了分子标记技术在水稻种质资源遗传多样性分析、重要功能基因的挖掘、品种DNA指纹库的构建及分子育种材料遗传基础的拓展等方面的作用,旨在为更充分的将分子标记技术应用于水稻种质资源的鉴定和分子育种提供参考。     

用RAPD和SSR分子标记鉴定小金海棠F1代 杂种实生苗的研究

小金海棠（Malus xiaojinensis）具有多种抗逆特性，是苹果属植物中一种珍贵的野生种质资源。由于小金海棠具有无融合生殖特性，早期准确、快速、有效地鉴定其F1代杂种实生苗对于研究利用其众多抗逆基因具有重要意义。RAPD技术是现有鉴定果树杂种实生苗最为简单、快速和有效的分子标记技术，但RAPD技术本身存在一定的不稳定性。笔者对小金海棠和山荆子（M. baccata）F1代杂交实生苗79个单株进行了RAPD分析，并用SSR标记对RAPD标记的初步鉴定结果进行了进一步的验证。结果显示，RAPD标记鉴定出的杂种实生苗有72.2%得到SSR标记的证实。研究的结果还表明，“叠加”和“新带”两种RAPD带型，结合RAPD标记特异位点数可以为小金海棠F1代杂种实生苗提供可靠的鉴定依据。研究为利用RAPD技术早期快速鉴定果树杂种实生苗提供了新的信息和依据。     

分子标记技术在甜菜育种中的应用

分子标记技术与常规育种技术相互紧密结合能显著提高育种效率。为了更好地阐明分子标记在甜菜育种中的作用,总结了国内外分子标记在甜菜亲缘关系及遗传多样性研究、遗传连锁图谱构建、数量性状基因定位（QTL）、分子标记辅助选择育种、杂种优势及种质鉴定中的研究现状和存在的问题。指出建立相应的高效分子标记辅助选育体系,创造出高产、优质、多抗或具广谱抗性的甜菜种质或品种是甜菜分子育种的研究方向。当前甜菜种质资源鉴定的关键任务是大力开发新型的分子标记进行甜菜种质资源遗传分析,绘制指纹图谱、进一步构建甜菜种质分子身份证。今后应加强对甜菜重要农艺性状基因进行精细定位,充分发掘QTL的信息,构建更为饱和的分子标记连锁图谱。     

RAPD分子标记技术在蔬菜研究中的应用

本文介绍了RAPD分子标记技术的原理和特点.综述了这种分子标记技术在蔬菜分类学及遗传多样性研究、遗传图谱构建、基因定位、分子标记辅助育种及品种纯度鉴定方面的应用情况,并对其应用前景进行了评述.     

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。     

家蚕抗核型多角体病毒病分子标记辅助育种

利用我国家蚕种质资源中发现的高抗NPV的材料NB和敏感材料306,组配近等基因系。采用RAPD技术获得分子标记,将标记转换成SCAR(sequence characterized amplified region)标记,利用SCAR标记开展了家蚕抗NPV新品种辅助育种选择,获得了家蚕抗NPV新品种。     

红花草莓红花基因RAPD标记转化为SCAR标记

红花草莓不但具有经济价值，还具有很高的观赏价值。本研究将3个与红花基因连锁的RAPD标记即AW65679（1031bp）、S484（620bp）与S1383（500bp）进行了克隆与核苷酸测序，并根据测序结果设计4对SCAR引物，将这4对引物对红花草莓品种粉红熊猫、白花草莓品种鬼怒甘及它们的杂交后代进行PCR扩增程序优化和鉴定，筛选出一对SCAR引物可扩增出与红花基因连锁的特异片段AW65679（1038bp），这就是与红花基因连锁的SCAR标记。SCAR标记因其稳定性好，重复性高将为草莓分子育种开辟一条新的有效途径。     

Identification of molecular markers associated with oleic and linolenic acid in spring oilseed rape (Brassica napus)

The quality of the oil derived from oilseed rape is determined by its fatty acid composition. Breeding oilseed rape for enhanced oil quality includes the development of cultivars with high oleic and low linolenic acid. Random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) techniques were investigated for the development of molecular markers for genes controlling oleic and/or linolenic acid. Markers that were identified were converted to sequence characterized amplified region (SCAR) markers for use in breeding. Molecular markers associated with these two fatty acids were identified in a doubled haploid population derived from a cross between the oilseed rape lines TO99‐5318‐20, very high oleic (>79%) and very low linolenic acid (<2%) × DH12075, high oleic (68%) and higher linolenic acid (>7%). Eight RAPD markers were associated with oleic and linolenic acid contents. The RAPD marker UBC 2₈₃₀ accounted for 43% and 13% of the genetic variation for oleic and linolenic acid levels, respectively. The RAPD marker UBC 153₅₅₀ accounted for 19% of the genetic variation for linolenic acid. The UBC 2₈₃₀ fragment was converted to a SCAR marker. The markers identified in this study should be useful tools for the early generation selection of high oleic and low linolenic acid genotypes in oilseed rape breeding programmes.     

Conversion of the RAPD OPC021150 marker of the Rfo restorer gene into a SCAR marker for rapid selection of oilseed rape

The Rfo fertility restorer gene for the Ogura cytoplasmic male sterility (CMS) applied for oilseed rape hybrid seed production can be monitored with the use of the RAPD OPC02₁₁₅₀ marker while molecular breeding. The aim of this work was to convert the RAPD marker into a more suitable SCAR marker. Total DNA was isolated from a doubled haploid line derived from the line BO20 (INRA, France). A fragment of 1150‐bp linked to the Rfo gene was PCR amplified with the use of the RAPD OPC02 primer, cloned and sequenced. A pair of primers was designed and PCR amplification was performed to develop a SCAR marker for the Rfo gene. The new marker was applied for analysis of 220 oilseed rape lines comprising doubled haploid and inbred restorer lines, restored hybrids as well as F₁ and F₂ recombinant generations involving restorer lines. Simultaneously, the RAPD OPC02 marker was used and it revealed that the markers are equivalent to each other. However, the developed new SCAR marker has made the analysis more practical, rapid and efficient.     

A SCAR marker for the sex types determination in Colombian genotypes of Carica papaya

Sex type determination in papaya (Carica papaya L.) is very important for crop improvement processes because it accelerates the identification of the fruitful plants. The use of molecular technology provides a quick and reliable identification of sex types in plantlets growing in seedbeds. Random amplified polymorphic DNA (RAPD) markers were used to determine the sex types of Colombian cultivars of dioecious papaya genotypes. This species has three sex types (male, female and hermaphrodite) determined by a multiallelic locus. There are no morphological differences at the chromosome level; therefore the identification of sex types by chromosomal dimorphism is not possible. A RAPD marker of 900 bp was found in male plants, but not in females or hermaphrodites. From this RAPD marker a sequence characterized amplified region (SCAR) was developed and it was possible to amplify fragments from the genomes of male and hermaphrodite plants, but not the female ones. The results indicate that this new SCAR marker will be valuable to determine the sex type of papaya plants.     

大葱雄性不育分子标记辅助选择的研究

大葱雄性不育在杂种优势利用中具有重要价值,传统方法选育不育系效率低,分子标记辅助育种可提高育种效率。本研究试图建立大葱雄性不育辅助育种的技术体系,加快大葱不育系、保持系的育种进程。利用RAPD技术分析了多态性片段S132800、S38960、S72300、S731100、S2002400与大葱育性的连锁关系,重组率分别为0、7．5％、0、4．2％、0。其中S132800、S2002400能在多数品种中区分N、S胞质,且重组率接近0,用于胞质鉴定具有很高的准确率,因而具有较高的利用价值。以S132800、S2002400为探针对不育系和保持系mtDNA的酶切片段进行了Southern杂交分析,结果表明它们在N、S胞质中存在多态性。预示着它们可能是与大葱CMS相关的片段。鉴于RAPD标记的应用有一定局限性,进一步对特异片段S132800、S2002400进行了克隆、测序和SCAR标记转化,其中S132800成功转化为SCAR标记,而S2002400转化后多态性消失。为进一步降低成本,简化了SCAR扩增产物的检测技术,初步建立了大葱不育系、保持系分子标记辅助选择的技术体系。研究表明,SCAR产物加入EB直接检测存在一定误差,而通过快速电泳可以快速、准确地鉴定单株的胞质类型。RAPD标记和SCAR标记在育种中应用有望提高大葱不育系、保持系的育种效率。     

普通菜豆抗炭疽病基因SCAR标记鉴定

利用12个菜豆品种（鉴别寄主）评价了7个抗炭疽病基因SCAR标记的可靠性和实用性,其中SBB14_1150/1050标记引物扩增没有特异性,SAS13₉₅₀没有扩增带。用5个可靠的菜豆抗炭疽病基因SCAR标记（SCAreoli₁₀₀₀、SH18₁₁₀₀、SAB3₄₀₀、SB12₃₅₀ 和SCF10₁₀₇₂）,对127份普通菜豆抗炭疽病品种进行抗炭疽病基因分子标记鉴定,82份未检测到SCAR标记,45份分别含有1~3个SCAR标记;检测到SCAR标记的资源中,13份含有SCAreoli₁₀₀₀标记,13份含有SH18₁₁₀₀标记,5份含有SAB3₄₀₀标记,9份含有SB12₃₅₀标记,11份含有SCF10₁₀₇₂标记。分析表明抗病品种含有的抗病基因标记与品种来源存在相关性。     

Conformity and genetic relatedness estimation in crop species having a narrow genetic base: the case of cucumber (Cucumis sativus L.)*

A set of 155 SSR (107) and SCAR (48) markers were used to evaluate 53 cucumber (Cucumis sativus L.) accessions of diverse origin to characterize genetic relationships and to define a standard marker array that was most effective in detecting genetic differences in this germplasm array. A multivariate marker‐based analysis of diverse germplasm using this standard marker array (17 SSR and 5 SCAR markers) was compared with results from a set of 70 previously reported RAPD markers, and then used to explore the potential value of these genetic markers for plant variety protection (PVP) and the establishment of essential derivation (ED) threshold values in this species using elite lines and hybrids and backcross progeny. Diversity analysis allowed identification of distinctly different lines that were used for the construction of three sets of backcross families (BC₁‐BC₃). While general genetic relationships among accessions were similar in SSR/SCAR analyses (r_s= 0.65) using two genetic distance (GD) estimators, differences in accession relationships were detected between RAPD and SSR/SCAR marker evaluations regardless of the estimator used. The GDs among elite germplasm with known pedigrees were relatively small (0.06‐0.23 for any pairwise comparison). GD values decreased and degree of fixation (at three to seven loci depending on the mating) increased with increased backcrossing such that recurrent parent allelic fixation occurred in least one family of each of the BC₃ families. In many instances the degree of fixation of loci was not uniformly achieved in the BC₃. Although the level of genetic polymorphisms will likely restrict the use of molecular markers for PVP and the establishment of ED values, the use of single nucleotide differences will likely provide opportunities to define specific functional distances that have potential for PVP in cucumber. Nevertheless, without an expanded, genetically robust standard marker array (e.g. 50 codominant markers), ED threshold values will be difficult to define in this species, and perhaps will require the appraisal of single nucleotide polymorphisms as discriminators of difference in this species.     

结球甘蓝迟抽薹基因RAPD标记转SCAR标记

本研究以与结球甘蓝迟抽薹基因连锁的N1750为引物,应用RAPD技术进行PCR扩增,检测到迟抽薹基因,对特异片段进行回收、克隆和测序,依据测序结果设计SCAR引物。在166株BC1群体(A21与P02杂交得到F1再与P02回交)中通过与RAPD标记的比较,SCAR扩增结果同RAPD扩增结果完全一致,从而证实了SCAR标记的准确性。实验结果表明,与甘蓝迟抽薹基因连锁的RAPD标记被成功转化为SCAR标记,为甘蓝分子标记辅助选择育种提供了基础。