猕猴桃性别鉴定研究进展

从形态、生理生化、同工酶、DNA分子标记等多方面综述了国内外在猕猴桃性别鉴定的研究进展.形态学、同工酶的猕猴桃性别鉴定方法相对简便但限制因素多,导致结果有时不理想;DNA分子标记较其它方法具有很多优点,其发展和应用给猕猴桃性别研究提供了一条有效的途径.现就同工酶、DNA分子标记在猕猴桃性别鉴定方面的应用进行了详细阐述,同时指出了它们在猕猴桃性别研究中存在的不足.     

AFLP标记在大白菜遗传育种中的应用

AFLP标记是一种新的DNA分子标记技术。对AFLP标记的基本原理、技术流程和关键技术做了简要介绍,从亲缘关系与遗传多样性分析、遗传图谱的构建、特定性状的连锁标记、品种纯度鉴定与QTL分析等方面概述了AFLP标记在大白菜遗传育种中的应用进展,并对其应用前景进行了展望。     

分子标记技术在西瓜甜瓜上的应用

近十几年来,AFLP、RAPD和SSR等分子标记技术的应用,加速了瓜类作物遗传和育种的步伐。根据有关文献介绍了分子标记技术在西瓜甜瓜遗传图谱构建、遗传多样性和物种亲缘关系、分子标记辅助选择、品种(系)指纹图谱构建及杂种纯度鉴定等研究领域的应用进展,其中,RAPD技术是一种高效的基因组DNA多态性分析技术,有快速、简便、高效等优点,主要综述了RAPD技术在西瓜、甜瓜遗传育种研究中的广泛应用,并探讨了该技术在使用中存在的问题及应用前景。     

SSR标记及其在葡萄上的应用

SSR(simple sequence repeat)作为第2代基于PCR的一种新型DNA分子遗传标记,数量丰富、多态性高、重复性好、分布于整个基因组、特异位点扩增、发生频率高、共显性遗传。对SSR引物开发,SSR标记在葡萄品种鉴别、系谱重建、遗传图谱构建、商业品种保护等方面的研究进展进行了综述,并指出了未来的研究方向。     

“十五”期间我国蔬菜分子育种研究进展

从分子遗传图谱的构建、遗传多样性分析,基因标记及定位,基因克隆及表达,杂种纯度鉴定与品种的DNA指纹分析以及基因工程育种等几个方面对“十五”期间我国蔬菜分子育种取得的进展进行了综述,对存在的问题进行了分析,并提出展望     

基于SRAP和SCoT标记的猕猴桃种质遗传多样性分析及变异材料鉴定

[目的]探明59份猕猴桃品种/种质的遗传背景及鉴定果肉全红型材料('H-16')的亲缘关系.[方法]利用SRAP及SCoT两种分子标记进行遗传多样性分析及变异鉴定.[结果]筛选出的12对SRAP引物和16条SCoT引物在59份材料中分别扩增出多态性条带125条和143条,平均多态性比率为99.07％和100％,平均遗传...     

AFLP技术在蔬菜作物遗传育种研究上的应用

综述了AFLP技术在蔬菜作物遗传图谱构建、遗传多样性和物种亲缘关系分析、重要基因的定位、基因的表达调控研究、蔬菜作物品种(系)的指纹图谱构建和杂种纯度鉴定、分子标记辅助选择等研究领域上的应用.并对该技术在蔬菜作物遗传育种研究上的应用前景进行了探讨.     

猕猴桃DNA条形码标记的筛选

以5种野生猕猴桃为试验材料,采用DNA条形码技术,对6个DNA条形码标记rbcL、matK、psbA、trnL-F、ITS、trnH-psbA进行序列分析,筛选出能鉴别猕猴桃种间分子差异的DNA条形码,以期利用DNA条形码技术用于猕猴桃育种。结果表明:通过序列碱基含量分析,trnL-F和ITS条形码序列的GC含量最高,为51.1%~55.2%,trnH-psbA的GC含量最小,约为32.0%;通过Blast比对,结果显示5个猕猴桃野生种与基因库中已登录种类的6个DNA条形码相似度在99%以上,而有些与基因库中已登录种类有所差异,体现出猕猴桃种内的遗传多样性或种间存在基因渗透;从系统进化树结果分析显示,ITS标记能将上述5个野生种明显区分,基因进化多样性两两比对分析显示,5个猕猴桃野生种之间差异明显;Tajima’s中性检验中,trnL-F,ITS和matK具有较高的核苷酸多样性和中性检验值。研究比较了6种DNA条形码标记,认为ITS种间多样性较高,差异明显,较适宜作为猕猴桃DNA条形码。     

分子标记技术在兰花遗传育种中的应用

分子标记技术在兰花遗传图谱构建、遗传多样性和物种亲缘关系、分子标记辅助选择、品种（系）指纹图谱构建及杂种纯度鉴定等研究领域可广泛应用.综述了随机扩增多态性DNA标记（RAPD）、扩增片段长度多态性标记（AFLP）、限制性片段长度多态性（RFLP）、单核苷酸多态性标记（SNP）等分子标记的基本原理、主要特点以及在兰花育种研究应用中的情况和展望.     

温郁金SCoT-PCR反应体系的正交优化

以温郁金为试材,运用L25(56)正交设计在5个水平上对影响温郁金SCoT-PCR反应的模板DNA、Mg2+、dNTPs、Taq酶和引物5个因素进行优化试验,对PCR结果进行极差分析。建立并优化温郁金的目标起始密码子多态性-聚合酶链式反应(SCoT-PCR)体系,以期为温郁金的遗传多样性分析及分子鉴定等研究提供技术支持。结果表明:建立了温郁金SCoT-PCR的最佳反应体系(20μL):引物0.8μmol/L,dNTPs 0.4mmol/L,Mg2+1.5mmol/L,Taq酶0.5U,模板DNA 40ng,且确定各因素对温郁金SCoT-PCR反应效果的影响大小依次为:dNTPsTaq酶引物Mg2+模板DNA,其中dNTPs对体系影响最大。优化的温郁金SCoT-PCR反应体系在多个温郁金品种遗传多样性研究中得到了验证,结果表现出良好的稳定性、重复性和多态性丰富等特点,可用于今后温郁金品种遗传多样性分析、系统发育分析、遗传图谱构建、基因定位和分子标记辅助育种等研究。     

基于EST-SSR标记分析猕猴桃种质遗传关系

利用开发的11对EST-SSR引物分析了33份猕猴桃种质资源的遗传多样性及其遗传关系。结果表明,11对EST-SSR引物在所有供试材料中均可扩增出清晰条带,其中有8对引物呈现多态性,多态性扩增率为72.7%,对33份种质材料的区分率达100%。8对多态性引物共检测到61个等位基因,每对引物可检测到的等位基因数为2-17,平均为7.6个。利用NTSYS-pc软件,以不加权成对算术平均法(UPGMA)对扩增结果进行聚类,谱系图显示,33份种质材料之间的相似系数在0.60～0.97,表明猕猴桃种质之间遗传关系相对来说不是很远。在相似系数0.73的水平上可将供试猕猴桃材料分为7个类群,其结果与传统的形态分类大体一致。从分子角度揭示了猕猴桃种质资源的遗传多态性及其遗传关系。可为猕猴桃种质改良提供理论依据。EST-SSR是非常有效和可靠的分子标记,可为猕猴桃分子育种及遗传连锁图构建奠定基础。     

Research advances on the genetic relationships of kiwifruit (Actinidia Lindl.)北大核心CSCD

Kiwifruit is an important fruit tree resource, and is one of the most successful fruit tree species for large-scale commercial cultivation worldwide. The natural distribution of kiwifruit is very extensive, but most of its taxa are mainly concentrated in the area south of the Qinling Mountains and east of the Hengduan Mountains. Chinese kiwifruit is not only a variety, but also different ecological habits. Chinese kiwifruit is rich in germplasm resources, contains a large number of wild resources, and has high genetic diversity, which can provide rich genetic basis and material conditions for the breeding of high- quality cultivars. Germplasm resources research has always been the most important foundation for fruit tree research, especially for breeding. The research on genetic relationship has become the core content of the research and utilization of fruit tree germplasm resources, which can provide a scientific basis for further exploration of species origin and evolution, systematic classification, germplasm conservation and utilization, and fruit tree breeding. Domestic and foreign scholars have studied the genetic relationship of kiwifruit germplasm resources from morphology, palynology, cytology, biochemical analysis to DNA molecular markers, from macroscopic level to microscopic level. The micromorphological characteristics, body size, cell structure and density of the leaf epidermal hair of the kiwifruit were-high polymorphism, which can be used as evidence for species identification. At present, the classification system of kiwifruit plants is mainly based on the type and degree of the indumentum of the fruit and leaves, the spots on the surface of the fruit and the morphology of the medulla of branches, and also giving consideration to the geographical distribution and other factors, as well as the combination of quantitative classification and computer processing methods. Liang C. F. divided the Chinese kiwifruit germplasms into four sections: Sect. Maculatae, Sect. Leiocarpae, Sect. Stellatae and Sect. Strigosae. The Sect. Leiocarpae can be divided into the Ser. Lamellatae and the Ser. Solidae, and the Sect. Stellatae can be divided into the Ser. Imperfectae and the Ser. Perfectae, which constitutes the current taxonomic system of four groups and four systems of Actinidia. However, the morphological characteristics of plants are susceptible to environmental conditions, and there are many subjective factors, which may lead to the divergence of taxonomic opinions. The morphological characteristics of kiwi pollen are controlled by genotypes, which are highly genetic conservation and are not affected by external conditions and carry a large amount of information. Related researchers used scanning electron microscopy to observe the pollen morphology of different species of kiwifruit. They found that the outer wall of the kiwi pollen has a large difference and has a significant polymorphism. They believe that the pollen grains of the genus have 3-colporate or 3-like colporate, which can be used as an important basis for packet classification. Kiwifruit has a wide range of chromosomal ploidy variations. Under natural conditions, diploid, tetraploid, hexaploid, octaploid and even dodecaploid occur, and the distribution pattern is reticulate the intraspecific and interspecific, as well as the sympatric distribution of multi species to various degrees. These make interspecific hybridization and gene introgression frequent, resulting in a series of rich genetic variations. Therefore, in-depth study on the interspecific and intraspecific genetic relationship of kiwifruit plants provides an important basis for breeding new varieties. The karyotype analysis of different ploidy revealed that the chromosome base of kiwifruit was 29, but there is still no clear conclusion about its chromosome origin. The research on the isozyme level shows that the kiwifruit has high genetic diversity at the level of cultivars and species, and there is a high degree of genetic heterozygosity and multiple alleles heterozygosity at its isozyme sites. At the DNA level, researchers used various methods of DNA molecular markers to detect different kiwi germplasm resources. The results showed that kiwifruit species or cultivars had rich genetic diversity, and there were high genetic diversity among different geographical regions. The genetic distance between kiwifruit cultivars was related to geographical distribution. In addition, studies on microsatellite showed that most kiwifruits showed higher heterozygosity, and polyploid kiwifruit had more genetic diversity. In addition to studying nuclear DNA, chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) have also received attention in kiwifruit research. The results show that the cpDNA of kiwifruit plants has strict paternal inheritance, and mtDNA has strict maternal inheritance. This rare genetic model complicates the genetic relationship between kiwifruit plants to a certain extent. In general, the research on the genetic relationship of kiwifruit has made great progress, especially the continuous improvement and mutual compensation of various molecular markers in recent years, making the research and analysis of genetic relationship more accurate and reliable. However, there are still many problems to be solved in the study of the genetic relationship of kiwifruit germplasm resources. In the future research, with the continuous innovation of research methods and the deepening of research content, the research on kiwifruit germplasm resources, especially the genetic relationship research, will surely achieve more results. It is of great scientific significance and application value for the rational utilization of abundant kiwifruit germplasm resources in China, especially the exploitation and utilization of a large number of excellent wild or semi-wild resources, and the breeding and industrial development of new kiwifruit varieties. © 2019 Journal of Fruit Science     

叶绿体DNA分析技术及其在栗属植物中的应用

被子植物叶绿体DNA母性遗传,有独立的进化路线,基于叶绿体DNA的分析技术在分子系统学、资源多样性评价、杂种鉴定等方面具有重要作用。栗属植物资源丰富,分布广泛。概述了叶绿体DNA分析技术在栗属植物中的研究进展,阐述了包括PCR-RFLP、DNA杂交、叶绿体SSR标记技术和叶绿体基因区段测序分析技术在内的分子标记技术的原理、特点及其在栗属植物中的应用。并对今后如何开发叶绿体DNA标记用于栗属植物的研究进行了展望。     

中华猕猴桃矮型性状EST-SSR连锁标记的筛选

以中华猕猴桃（Actinidia chinensis Planch）矮型种质‘赣猕5号’为母本,普通型中华猕猴桃‘奉雄2号’为父本构建F1群体,采用优越而高效的基于表达序列标签（Expressed Sequence Tags,EST）的简单序列重复（Simple Sequence Repeats,SSR）标记技术,结合群体分离分析法（Bulked Segregant Analysis,BSA）在荧光DNA测序仪（ABI3130）上进行了‘赣猕5号’矮型性状连锁的EST-SSR分子标记研究。结果表明,通过本实验室开发并设计的85对EST-SSR荧光引物的扩增筛选,其中引物EST-Ad042在亲本及其矮型与普通型DNA 池之间扩增出一条285 bp 的多态性片段,经对其F1代分离群体部分矮型与普通型单株的随机验证,该片段稳定出现,在矮型植株中表现为有带,在普通型植株中表现为无带。遗传连锁分析表明,该标记与矮型基因之间的连锁距离为8.8 cM。EST-SSR荧光引物能够有效地筛选到中华猕猴桃的特异标记,可以作为鉴别矮型与普通型植株的标记引物。     

ISSR及其在果树上的应用

ISSR是一种基于微卫星序列发展起来的新的分子标记,具有简便、稳定、DNA多态性高等优点。ISSR标记呈孟德尔式遗传,大部分ISSR标记为显性标记。综合有关文献就ISSR的原理、操作及其在果树种质鉴定、遗传多样性检测、亲缘关系分析和遗传图谱构建等方面的应用和进展进行简要的阐述。     

野生毛花猕猴桃果实表型性状及SSR遗传多样性分析

以江西省武功山境内的70份野生毛花猕猴桃种质资源为试材,对其果实表型性状、SSR遗传多样性及其亲缘关系进行分析和评价。根据UPOV（国际植物新品种保护联盟）公布的猕猴桃属品种测定标准对供试材料的果实表型性状进行观测。参照猕猴桃遗传连锁图谱,选用分布于中华猕猴桃基因组中的70对SSR引物对供试材料进行多态性分析。结果表明,在70对引物中,21对引物成功扩增出多态性片段。随机采集的70份野生毛花猕猴桃种质资源中,其果实表型性状和DNA分子水平均存在丰富的遗传多样性。基于UPGMA聚类分析将供试的野生毛花猕猴桃资源分为果实圆形和椭圆形混合组、果实椭圆形组以及果实圆柱形组。21对多态性高的SSR引物共检测出127个等位变异位点,变异范围在2 ~ 12之间,平均每对SSR引物可检测到6.04个等位位点。遗传相似系数（GS）变异范围为0.5306 ~ 0.9252。GS值在0.65水平上UPGMA聚类分析可将供试的野生毛花猕猴桃种质资源划分成Ⅰ、Ⅱ和 Ⅲ 共3个组,这与果实表型性状分析的结果基本一致。这些遗传变异丰富的种质资源可为猕猴桃育种提供有价值的材料。     

ISSR和SRAP标记技术在兰花植物种质资源研究中的应用

分子标记技术在兰花种质资源的亲缘关系鉴定与遗传多样性分析、分子遗传图谱的构建与目的性状基因的标记定位、基因库的构建和基因克隆、辅助育种选择及品种纯度鉴定等研究领域被广泛利用与推广.现从简单重复序列间扩增(Inter-simple sequence repeat,ISSR)和相关序列扩增多态性(SRAP)分子标记技术的基本原理、方法、特点以及在兰花植物种质资源的遗传育种中的应用现状和前景进行了综述与探讨.     

豆瓣菜种质资源遗传多样性的RAPD和ISSR分析

以8个豆瓣菜的品种为试材，用筛选出的79个RAPD引物和34个ISSR引物对这8个品种的基因组DNA进行扩增，分别扩增出361条和179条谱带，每个引物扩增出的带在3～10条之间，平均每个引物扩增出约5条带。根据所得的条带进行聚类分析，两种标记产生的聚类图存在一些差异，但它们都可以较好地将8个品种按亲缘关系的远近划分为3个不同的类群。Mantel测试得出相关系数r=0．58155，表明RAPD和ISSR两种分子标记技术的相关度很低。     

分子标记技术在莲遗传多样性分析中的应用

分子标记是继形态标记、细胞标记、生化标记之后出现的一种全新的遗传标记方法,已被广泛地应用于生命科学研究的各个领域。主要介绍了现阶段常用的几种分子标记技术,以及这些标记技术在莲遗传多样性分析中的应用,并对其应用前景进行了展望。